Colorectal cancer detection kit or device, and detection method

ABSTRACT

It is intended to provide a kit or a device for the detection of colorectal cancer and a method for detecting colorectal cancer. The present invention provides a kit or a device for the detection of colorectal cancer, comprising a nucleic acid capable of specifically binding to a miRNA in a sample from a subject, and a method for detecting colorectal cancer, comprising measuring the miRNA in vitro.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a Divisional of U.S. application Ser. No. 16/789,986filed Feb. 13, 2020, which is a Divisional of application Ser. No.15/318,312, filed on Dec. 12, 2016 (now U.S. Pat. No. 10,604,810), whichis the National Phase under 35 U.S.C. § 371 of International ApplicationNo. PCT/JP2015/066970, filed on Jun. 12, 2015, which claims the benefitunder 35 U.S.C. § 119(a) to Patent Application No. 2014-122686, filed inJapan on Jun. 13, 2014, and to Patent Application No. 2015-070182, filedin Japan on Mar. 30, 2015, all of which are hereby expresslyincorporated by reference into the present application.

REFERENCE TO ELECTRONIC SEQUENCE LISTING

The application contains a Sequence Listing which has been submittedelectronically in .XML format and is hereby incorporated by reference inits entirety. Said .XML copy, created on Sep. 12, 2022, is named“PH-6235-PCT-US-DIV1-DIV1 Sequence Listing ST26” and is 571,313 bytes insize. The sequence listing contained in this .XML file is part of thespecification and is hereby incorporated by reference herein in itsentirety.

TECHNICAL FIELD

The present invention relates to a kit or a device for the detection ofcolorectal cancer, comprising a nucleic acid capable of specificallybinding to a particular miRNA, which is used for examining the presenceor absence of colorectal cancer in a subject, and a method for detectingcolorectal cancer, comprising measuring an expression level of the miRNAusing the nucleic acid.

BACKGROUND ART

The large intestine is an organ that stores residual bowel contentsafter digestion and absorption, and produces feces while absorbingwater. The large intestine begins with the cecum, which is thenconnected to the ascending colon, the transverse colon, the descendingcolon, the sigmoid colon, the rectum, and the anal canal. According tothe 2011 type-specific cancer statistics in Japan disclosed by theCenter for Cancer Control and Information Services, National CancerCenter, the number of individuals affected by colorectal cancer was112,772 people. Namely, it is estimated that one in approximately 14Japanese people experience colorectal cancer. The number of incidencesof this cancer takes the 2nd place by cancer site. The number ofcolorectal cancer deaths in men and women together climbs to 45,744people and takes the 3rd place by cancer site. It is estimated that onein approximately 20 Americans develop colorectal cancer. The estimatednumber of American individuals affected by colorectal cancer climbed to96,830 people in 2014, among which approximately 40,000 peoplereportedly died (Non Patent Literature 1).

The progression stages of colorectal cancer are specified in Non PatentLiterature 2 and classified into stage 0 (Tis/N0/M0), stage I (T1 toT2/N0/M0), stage II (T3 to T4/N0/M0), stage IIA (T3/N0/M0), stage IIB(T4a/N0/M0), stage IIC (T4b/N0/M0), stage III (N1 to N2/M0), stage IIIA(T1 to T2/N1/M0 and T1/N2a/M0), stage IIIB (T3 to T4a/N1/M0 and T2 toT3/N2a/M0 and T1 to T2/N2b/M0), stage IIIC (T4a/N2a/M0 and T3 toT4a/N2b/M0 and T4b/N1 to N2/M0), stage IVA (M1a), and stage IVB (M1b)according to the degrees of tumor spread (Tis and T1 to T4), lymph nodemetastasis (N0, N1a to N1c, and N2a to N2b), and distant metastasis (M0and M1a to M1b).

The survival rate of colorectal cancer differs depending on the stagesof progression. Non Patent Literature 1 has reported the followingrespective statistic values of colon cancer and rectal cancer. The5-year relative survival rate of colon cancer is reportedly 74% forstage I, 67% for stage IIA, 59% for stage IIB, 37% for stage IIC, 73%for stage IIIA, 46% for stage IIIB, 28% for stage IIIC, and 6% for stageIV. Also, the 5-year relative survival rate of rectal cancer isreportedly 74% for stage I, 65% for stage IIA, 52% for stage IIB, 32%for stage IIC, 74% for stage IIIA, 45% for stage IIIB, 33% for stageIIIC, and 6% for stage IV. Evidently, colorectal cancer at an earlystage of progression leads to a high survival rate. Thus, the earlydetection and treatment of colorectal cancer makes a significantcontribution to improvement in survival rate.

The treatment of colorectal cancer is mainly laparotomy or laparoscopicsurgery, which is often used in combination with postoperativeanticancer drug treatment or radiotherapy (Non Patent Literature 1).Particularly, early colorectal cancer may be adaptable to endoscopicsurgery which permits treatment without abdominal resection.

As described in Non Patent Literature 1, fecal occult blood test andendoscopy are widely prevalent as tests of colorectal cancer.Particularly, the fecal occult blood test is inexpensive and noninvasiveand is also carried out at home. Therefore, the American Cancer Societyrecommends taking the fecal occult blood test every year. In order tofurther examine a tumor site and spread of the cancer, an imaging testsuch as barium enema, CT, or MRI is also carried out in addition to thecolonoscopy. Alternatively, tests on blood tumor markers such as CEA andCA19-9 may be carried out for the purpose of observing the prognosis orthe therapeutic effects on patients already diagnosed with colorectalcancer (Non Patent Literature 1).

As shown in Patent Literatures 1 to 4, there are reports, albeit at aresearch stage, on the detection of colorectal cancer using theexpression levels of microRNAs (miRNAs) or combinations of theexpression levels of miRNAs and the expression levels of additionalprotein markers in biological samples including blood.

Patent Literature 1 discloses a method for detecting colorectal canceror other cancers using hsa-miR-92a-2-5p, hsa-miR-128-2-5p, andhsa-miR-24-3p in colorectal cancer tissues.

Patent Literature 2 discloses a method for detecting colorectal cancerusing hsa-miR-1233-5p and hsa-miR-1225-3p in plasma.

Patent Literature 3 discloses a method for detecting colorectal cancerusing multiple miRNAs such as hsa-miR-1231, hsa-miR-423-5p, andhsa-miR-1268a in large intestine tissues or feces.

Patent Literature 4 discloses a method for detecting colorectal cancerusing hsa-miR-150-3p, miR-92a-2-5p, and the like in tissues.

CITATION LIST Patent Literature

-   Patent Literature 1: International Publication No. WO 2007/081740-   Patent Literature 2: U.S. Patent Application Publication No.    2013/102487-   Patent Literature 3: U.S. Patent Application Publication No.    2012/088687-   Patent Literature 4: JP Patent Publication (Kohyo) No. 2009-531019 A    (2009)

Non Patent Literature

-   Non Patent Literature 1: American Cancer Society, “Colorectal    Cancer”, 2013, p. 5 to 6, 17 to 28, 33 to, 45 to 54, and 67 to 71-   Non Patent Literature 2: Sobin, L. et al., “TNM Classification of    Malignant Tumours, the 7th edition”, 2010, p. 94-99-   Non Patent Literature 3: Allison, J E. et al., 1996, The New England    Journal of Medicine, Vol. 334 (3), p. 155-9-   Non Patent Literature 4: Palmqvist, R. et al., 2007, Diseases of    colon and rectum, Vol. 46 (11), p. 1538-44

SUMMARY OF INVENTION Technical Problem

An object of the present invention is to find a novel tumor marker forcolorectal cancer and to provide a method that can effectively detectcolorectal cancer using a nucleic acid capable of specifically bindingto the marker. The fecal occult blood test, which is widely used atpresent as a first test of colorectal cancer, produces positive resultseven for non-cancerous reasons such as hemorrhoid, whereas this testfails to detect early colorectal cancer without bleeding and overlooks90% or more of abnormalities in the large intestine (including cancer)according to the report (Non Patent Literature 1). The specificsensitivity of the fecal occult blood test differs largely from 37% to79.4% depending on a testing kit used, and its specificity is reportedly86.7% to 97.7% (Non Patent Literature 3). Although the colonoscopy isknown to have high examination accuracy, this examination is difficultto apply as a primary screening because of the necessity of pretreatmentor sedatives on patients, relatively high cost, etc. (Non PatentLiterature 1). The tumor markers such as CEA and CA19-9 in blood mayelevate in cancers other than colorectal cancer and therefore allegedlyfail to determine the presence or absence of colorectal cancer. Thefalse diagnosis of other cancers as colorectal cancer wastes appropriatetherapeutic opportunity or places unnecessary economical and physicalburdens on patients due to the application of wrong medicine. Therefore,use of CEA or CA19-9 is often limited to the observation of theprognosis and of therapeutic effects on patients already diagnosed withcolorectal cancer (Non Patent Literature 1). The report states that theCEA test has specificity of 99%, but sensitivity of only 12%, suggestingthat the significance of tumor marker measurement as a colorectal cancerscreening test is poor (Non Patent Literature 4).

As described below, there are reports, albeit at a research stage, onthe determination of colorectal cancer using the expression levels ofmicroRNAs (miRNAs) in biological samples including blood, none of which,however, have yet been brought into practical use.

Patent Literature 1 discloses a method for detecting colorectal canceror other cancers using hsa-miR-92a-2-5p, hsa-miR-128-2-5p, andhsa-miR-24-3p in colorectal cancer tissues. This detection method,however, requires obtaining colorectal cancer tissue samples by surgicaloperation, and this step places a heavy physical burden on patients.Therefore, this method is not favorable as an examination method. Inaddition, this detection method does not describe specific colorectalcancer detection performance such as accuracy, sensitivity, orspecificity and is thus industrially less practical.

Patent Literature 3 discloses a method for detecting colorectal cancerusing multiple miRNAs such as hsa-miR-1231, hsa-miR-423-5p, andhsa-miR-1268a in large intestine tissues or feces. Since surgicaloperation for obtaining colorectal cancer tissues places a heavyphysical burden on patients, this method is not favorable as anexamination method. In addition, although the collection of fecalsamples is noninvasive, test substances may exist unevenly in feces.This tends to cause unfavorable variations in testing results.

Patent Literature 4 discloses a method for detecting colorectal cancerusing hsa-miR-150-3p, miR-92a-2-5p, and the like in tissues. Thisliterature, however, neither describes detection performance such asaccuracy, sensitivity, or specificity nor describes a specific methodfor determining colorectal cancer using blood. Therefore, this method isindustrially less practical. In addition, these miRNA markers were notvalidated in an independent sample group and are thus less reliable.

As mentioned above, the existing tumor markers exhibit low performancein the detection of colorectal cancer, or neither performance nordetection methods are specifically shown as to the markers at a researchstage. Therefore, use of these markers might lead to imposing needlessextra examination due to the false detection of healthy subjects asbeing colorectal cancer patients, or might waste therapeutic opportunitybecause of overlooking colorectal cancer patients. In addition, themeasurement of several dozens to several hundreds of miRNAs increasesexamination cost and is therefore difficult to use in large-scalescreening such as medical checkup. Furthermore, the collection ofcolorectal tissues for measuring the tumor markers is highly invasive topatients and is not favorable. Hence, there is a demand for a highlyaccurate colorectal cancer marker that is detectable from blood, whichcan be collected in a less invasive manner, and is capable of correctlydetermining a colorectal cancer patient as a colorectal cancer patientand a healthy subject as a healthy subject. The early detection andtreatment of colorectal cancer can drastically improve survival rates.Furthermore, the early detection of colorectal cancer leads to theapplicability of endoscopic surgery which permits treatment withoutabdominal resection. Therefore, a highly sensitive colorectal cancermarker that can detect colorectal cancer even at an early stage ofprogression is desired.

Solution to Problem

The present inventors have conducted diligent studies to attain theobject and consequently completed the present invention by findingmultiple genes usable as markers for the detection of colorectal cancerfrom blood, which can be collected with limited invasiveness, andfinding that colorectal cancer can be significantly detected by using anucleic acid capable of specifically binding to any of these markers.

SUMMARY OF INVENTION

Specifically, the present invention has the following features:

(1) A kit for the detection of colorectal cancer, comprising a nucleicacid capable of specifically binding to at least one or morepolynucleotide(s) selected from the group consisting of colorectalcancer markers miR-6726-5p, miR-4257, miR-6787-5p, miR-6780b-5p,miR-3131, miR-7108-5p, miR-1343-3p, miR-1247-3p, miR-4651, miR-6757-5p,miR-3679-5p, miR-7641, miR-6746-5p, miR-8072, miR-6741-5p, miR-1908-5p,miR-6857-5p, miR-4746-3p, miR-744-5p, miR-4792, miR-564, miR-6791-5p,miR-6825-5p, miR-6826-5p, miR-4665-3p, miR-4467, miR-3188, miR-6125,miR-6756-5p, miR-1228-3p, miR-8063, miR-8069, miR-6875-5p, miR-3185,miR-4433b-3p, miR-6887-5p, miR-128-1-5p, miR-6724-5p, miR-1914-3p,miR-1225-5p, miR-4419b, miR-7110-5p, miR-187-5p, miR-3184-5p,miR-204-3p, miR-5572, miR-6729-5p, miR-615-5p, miR-6749-5p, miR-6515-3p,miR-3937, miR-6840-3p, miR-6893-5p, miR-4728-5p, miR-6717-5p,miR-7113-3p, miR-4665-5p, miR-642b-3p, miR-7109-5p, miR-6842-5p,miR-4442, miR-4433-3p, miR-4707-5p, miR-6126, miR-4449, miR-4706,miR-1913, miR-602, miR-939-5p, miR-4695-5p, miR-711, miR-6816-5p,miR-4632-5p, miR-6721-5p, miR-7847-3p, miR-6132, miR-887-3p,miR-3679-3p, miR-6784-5p, miR-1249, miR-937-5p, miR-5195-3p,miR-6732-5p, miR-4417, miR-4281, miR-4734, miR-6766-3p, miR-663a,miR-4513, miR-6781-5p, miR-1227-5p, miR-6845-5p, miR-6798-5p,miR-3620-5p, miR-1915-5p, miR-4294, miR-642a-3p, miR-371a-5p, miR-940,miR-4450, miR-4723-5p, miR-1469, miR-6861-5p, miR-7975, miR-6879-5p,miR-6802-5p, miR-1268b, miR-663b, miR-125a-3p, miR-2861, miR-6088,miR-4758-5p, miR-296-3p, miR-6738-5p, miR-671-5p, miR-4454, miR-4516,miR-7845-5p, miR-4741, miR-92b-5p, miR-6795-5p, miR-6805-3p,miR-4725-3p, miR-6782-5p, miR-4688, miR-6850-5p, miR-6777-5p,miR-6785-5p, miR-7106-5p, miR-3663-3p, miR-6131, miR-1915-3p, miR-4532,miR-6820-5p, miR-4689, miR-4638-5p, miR-3656, miR-3621, miR-6769b-5p,miR-149-3p, miR-23b-3p, miR-3135b, miR-6848-5p, miR-6769a-5p, miR-4327,miR-6765-3p, miR-6716-5p, miR-6877-5p, miR-6727-5p, miR-4534, miR-614,miR-1202, miR-575, miR-6870-5p, miR-6722-3p, miR-7977, miR-4649-5p,miR-4675, miR-6075, miR-6779-5p, miR-4271, miR-3196, miR-6803-5p,miR-6789-5p, miR-4648, miR-4508, miR-4749-5p, miR-4505, miR-5698,miR-1199-5p, miR-4763-3p, miR-6836-3p, miR-3195, miR-718, miR-3178,miR-638, miR-4497, miR-6085, miR-6752-5p and miR-135a-3p.

(2) The kit according to (1), wherein miR-6726-5p is hsa-miR-6726-5p,miR-4257 is hsa-miR-4257, miR-6787-5p is hsa-miR-6787-5p, miR-6780b-5pis hsa-miR-6780b-5p, miR-3131 is hsa-miR-3131, miR-7108-5p ishsa-miR-7108-5p, miR-1343-3p is hsa-miR-1343-3p, miR-1247-3p ishsa-miR-1247-3p, miR-4651 is hsa-miR-4651, miR-6757-5p ishsa-miR-6757-5p, miR-3679-5p is hsa-miR-3679-5p, miR-7641 ishsa-miR-7641, miR-6746-5p is hsa-miR-6746-5p, miR-8072 is hsa-miR-8072,miR-6741-5p is hsa-miR-6741-5p, miR-1908-5p is hsa-miR-1908-5p,miR-6857-5p is hsa-miR-6857-5p, miR-4746-3p is hsa-miR-4746-3p,miR-744-5p is hsa-miR-744-5p, miR-4792 is hsa-miR-4792, miR-564 ishsa-miR-564, miR-6791-5p is hsa-miR-6791-5p, miR-6825-5p ishsa-miR-6825-5p, miR-6826-5p is hsa-miR-6826-5p, miR-4665-3p ishsa-miR-4665-3p, miR-4467 is hsa-miR-4467, miR-3188 is hsa-miR-3188,miR-6125 is hsa-miR-6125, miR-6756-5p is hsa-miR-6756-5p, miR-1228-3p ishsa-miR-1228-3p, miR-8063 is hsa-miR-8063, miR-8069 is hsa-miR-8069,miR-6875-5p is hsa-miR-6875-5p, miR-3185 is hsa-miR-3185, miR-4433b-3pis hsa-miR-4433b-3p, miR-6887-5p is hsa-miR-6887-5p, miR-128-1-5p ishsa-miR-128-1-5p, miR-6724-5p is hsa-miR-6724-5p, miR-1914-3p ishsa-miR-1914-3p, miR-1225-5p is hsa-miR-1225-5p, miR-4419b ishsa-miR-4419b, miR-7110-5p is hsa-miR-7110-5p, miR-187-5p ishsa-miR-187-5p, miR-3184-5p is hsa-miR-3184-5p, miR-204-3p ishsa-miR-204-3p, miR-5572 is hsa-miR-5572, miR-6729-5p ishsa-miR-6729-5p, miR-615-5p is hsa-miR-615-5p, miR-6749-5p ishsa-miR-6749-5p, miR-6515-3p is hsa-miR-6515-3p, miR-3937 ishsa-miR-3937, miR-6840-3p is hsa-miR-6840-3p, miR-6893-5p ishsa-miR-6893-5p, miR-4728-5p is hsa-miR-4728-5p, miR-6717-5p ishsa-miR-6717-5p, miR-7113-3p is hsa-miR-7113-3p, miR-4665-5p ishsa-miR-4665-5p, miR-642b-3p is hsa-miR-642b-3p, miR-7109-5p ishsa-miR-7109-5p, miR-6842-5p is hsa-miR-6842-5p, miR-4442 ishsa-miR-4442, miR-4433-3p is hsa-miR-4433-3p, miR-4707-5p ishsa-miR-4707-5p, miR-6126 is hsa-miR-6126, miR-4449 is hsa-miR-4449,miR-4706 is hsa-miR-4706, miR-1913 is hsa-miR-1913, miR-602 ishsa-miR-602, miR-939-5p is hsa-miR-939-5p, miR-4695-5p ishsa-miR-4695-5p, miR-711 is hsa-miR-711, miR-6816-5p is hsa-miR-6816-5p,miR-4632-5p is hsa-miR-4632-5p, miR-6721-5p is hsa-miR-6721-5p,miR-7847-3p is hsa-miR-7847-3p, miR-6132 is hsa-miR-6132, miR-887-3p ishsa-miR-887-3p, miR-3679-3p is hsa-miR-3679-3p, miR-6784-5p ishsa-miR-6784-5p, miR-1249 is hsa-miR-1249, miR-937-5p is hsa-miR-937-5p,miR-5195-3p is hsa-miR-5195-3p, miR-6732-5p is hsa-miR-6732-5p, miR-4417is hsa-miR-4417, miR-4281 is hsa-miR-4281, miR-4734 is hsa-miR-4734,miR-6766-3p is hsa-miR-6766-3p, miR-663a is hsa-miR-663a, miR-4513 ishsa-miR-4513, miR-6781-5p is hsa-miR-6781-5p, miR-1227-5p ishsa-miR-1227-5p, miR-6845-5p is hsa-miR-6845-5p, miR-6798-5p ishsa-miR-6798-5p, miR-3620-5p is hsa-miR-3620-5p, miR-1915-5p ishsa-miR-1915-5p, miR-4294 is hsa-miR-4294, miR-642a-3p ishsa-miR-642a-3p, miR-371a-5p is hsa-miR-371a-5p, miR-940 is hsa-miR-940,miR-4450 is hsa-miR-4450, miR-4723-5p is hsa-miR-4723-5p, miR-1469 ishsa-miR-1469, miR-6861-5p is hsa-miR-6861-5p, miR-7975 is hsa-miR-7975,miR-6879-5p is hsa-miR-6879-5p, miR-6802-5p is hsa-miR-6802-5p,miR-1268b is hsa-miR-1268b, miR-663b is hsa-miR-663b, miR-125a-3p ishsa-miR-125a-3p, miR-2861 is hsa-miR-2861, miR-6088 is hsa-miR-6088,miR-4758-5p is hsa-miR-4758-5p, miR-296-3p is hsa-miR-296-3p,miR-6738-5p is hsa-miR-6738-5p, miR-671-5p is hsa-miR-671-5p, miR-4454is hsa-miR-4454, miR-4516 is hsa-miR-4516, miR-7845-5p ishsa-miR-7845-5p, miR-4741 is hsa-miR-4741, miR-92b-5p is hsa-miR-92b-5p,miR-6795-5p is hsa-miR-6795-5p, miR-6805-3p is hsa-miR-6805-3p,miR-4725-3p is hsa-miR-4725-3p, miR-6782-5p is hsa-miR-6782-5p, miR-4688is hsa-miR-4688, miR-6850-5p is hsa-miR-6850-5p, miR-6777-5p ishsa-miR-6777-5p, miR-6785-5p is hsa-miR-6785-5p, miR-7106-5p ishsa-miR-7106-5p, miR-3663-3p is hsa-miR-3663-3p, miR-6131 ishsa-miR-6131, miR-1915-3p is hsa-miR-1915-3p, miR-4532 is hsa-miR-4532,miR-6820-5p is hsa-miR-6820-5p, miR-4689 is hsa-miR-4689, miR-4638-5p ishsa-miR-4638-5p, miR-3656 is hsa-miR-3656, miR-3621 is hsa-miR-3621,miR-6769b-5p is hsa-miR-6769b-5p, miR-149-3p is hsa-miR-149-3p,miR-23b-3p is hsa-miR-23b-3p, miR-3135b is hsa-miR-3135b, miR-6848-5p ishsa-miR-6848-5p, miR-6769a-5p is hsa-miR-6769a-5p, miR-4327 ishsa-miR-4327, miR-6765-3p is hsa-miR-6765-3p, miR-6716-5p ishsa-miR-6716-5p, miR-6877-5p is hsa-miR-6877-5p, miR-6727-5p ishsa-miR-6727-5p, miR-4534 is hsa-miR-4534, miR-614 is hsa-miR-614,miR-1202 is hsa-miR-1202, miR-575 is hsa-miR-575, miR-6870-5p ishsa-miR-6870-5p, miR-6722-3p is hsa-miR-6722-3p, miR-7977 ishsa-miR-7977, miR-4649-5p is hsa-miR-4649-5p, miR-4675 is hsa-miR-4675,miR-6075 is hsa-miR-6075, miR-6779-5p is hsa-miR-6779-5p, miR-4271 ishsa-miR-4271, miR-3196 is hsa-miR-3196, miR-6803-5p is hsa-miR-6803-5p,miR-6789-5p is hsa-miR-6789-5p, miR-4648 is hsa-miR-4648, miR-4508 ishsa-miR-4508, miR-4749-5p is hsa-miR-4749-5p, miR-4505 is hsa-miR-4505,miR-5698 is hsa-miR-5698, miR-1199-5p is hsa-miR-1199-5p, miR-4763-3p ishsa-miR-4763-3p, miR-6836-3p is hsa-miR-6836-3p, miR-3195 ishsa-miR-3195, miR-718 is hsa-miR-718, miR-3178 is hsa-miR-3178, miR-638is hsa-miR-638, miR-4497 is hsa-miR-4497, miR-6085 is hsa-miR-6085,miR-6752-5p is hsa-miR-6752-5p, and miR-135a-3p is hsa-miR-135a-3p.

(3) The kit according to (1) or (2), wherein the nucleic acid is apolynucleotide selected from the group consisting of the followingpolynucleotides (a) to (e):

(a) a polynucleotide consisting of a nucleotide sequence represented byany of SEQ ID NOs: 1 to 171 and 606 to 614 or a nucleotide sequencederived from the nucleotide sequence by the replacement of u with t, avariant thereof, a derivative thereof, or a fragment thereof comprising15 or more consecutive nucleotides,(b) a polynucleotide comprising a nucleotide sequence represented by anyof SEQ ID NOs: 1 to 171 and 606 to 614,(c) a polynucleotide consisting of a nucleotide sequence complementaryto a nucleotide sequence represented by any of SEQ ID NOs: 1 to 171 and606 to 614 or a nucleotide sequence derived from the nucleotide sequenceby the replacement of u with t, a variant thereof, a derivative thereof,or a fragment thereof comprising 15 or more consecutive nucleotides,(d) a polynucleotide comprising a nucleotide sequence complementary to anucleotide sequence represented by any of SEQ ID NOs: 1 to 171 and 606to 614 or a nucleotide sequence derived from the nucleotide sequence bythe replacement of u with t, and(e) a polynucleotide hybridizing under stringent conditions to any ofthe polynucleotides (a) to (d).

(4) The kit according to any of (1) to (3), wherein the kit furthercomprises a nucleic acid capable of specifically binding to at least oneor more polynucleotide(s) selected from the group consisting of othercolorectal cancer markers miR-1231-5p, miR-1233-5p, miR-150-3p,miR-1225-3p, miR-92a-2-5p, miR-423-5p, miR-1268a, miR-128-2-5p andmiR-24-3p.

(5) The kit according to (4), wherein miR-1231 is hsa-miR-1231,miR-1233-5p is hsa-miR-1233-5p, miR-150-3p is hsa-miR-150-3p,miR-1225-3p is hsa-miR-1225-3p, miR-92a-2-5p is hsa-miR-92a-2-5p,miR-423-5p is hsa-miR-423-5p, miR-1268a is hsa-miR-1268a, miR-128-2-5pis hsa-miR-128-2-5p, and miR-24-3p is hsa-miR-24-3p.

(6) The kit according to (4) or (5), wherein the nucleic acid is apolynucleotide selected from the group consisting of the followingpolynucleotides (f) to (j):

(f) a polynucleotide consisting of a nucleotide sequence represented byany of SEQ ID NOs: 172 to 180 or a nucleotide sequence derived from thenucleotide sequence by the replacement of u with t, a variant thereof, aderivative thereof, or a fragment thereof comprising 15 or moreconsecutive nucleotides,(g) a polynucleotide comprising a nucleotide sequence represented by anyof SEQ ID NOs: 172 to 180,(h) a polynucleotide consisting of a nucleotide sequence complementaryto a nucleotide sequence represented by any of SEQ ID NOs: 172 to 180 ora nucleotide sequence derived from the nucleotide sequence by thereplacement of u with t, a variant thereof, a derivative thereof, or afragment thereof comprising 15 or more consecutive nucleotides,(i) a polynucleotide comprising a nucleotide sequence complementary to anucleotide sequence represented by any of SEQ ID NOs: 172 to 180 or anucleotide sequence derived from the nucleotide sequence by thereplacement of u with t, and(j) a polynucleotide hybridizing under stringent conditions to any ofthe polynucleotides (f) to (i).

(7) The kit according to any of (1) to (6), wherein the kit furthercomprises a nucleic acid capable of specifically binding to at least oneor more polynucleotide(s) selected from the group consisting of othercolorectal cancer markers miR-4697-5p, miR-3197, miR-675-5p, miR-4486,miR-7107-5p, miR-23a-3p, miR-4667-5p, miR-451a, miR-3940-5p, miR-8059,miR-6813-5p, miR-4492, miR-4476 and miR-6090.

(8) The kit according to (7), wherein miR-4697-5p is hsa-miR-4697-5p,miR-3197 is hsa-miR-3197, miR-675-5p is hsa-miR-675-5p, miR-4486 ishsa-miR-4486, miR-7107-5p is hsa-miR-7107-5p, miR-23a-3p ishsa-miR-23a-3p, miR-4667-5p is hsa-miR-4667-5p, miR-451a ishsa-miR-451a, miR-3940-5p is hsa-miR-3940-5p, miR-8059 is hsa-miR-8059,miR-6813-5p is hsa-miR-6813-5p, miR-4492 is hsa-miR-4492, miR-4476 ishsa-miR-4476, and miR-6090 is hsa-miR-6090.

(9) The kit according to (7) or (8), wherein the nucleic acid is apolynucleotide selected from the group consisting of the followingpolynucleotides (k) to (o):

(k) a polynucleotide consisting of a nucleotide sequence represented byany of SEQ ID NOs: 181 to 194 or a nucleotide sequence derived from thenucleotide sequence by the replacement of u with t, a variant thereof, aderivative thereof, or a fragment thereof comprising 15 or moreconsecutive nucleotides,(l) a polynucleotide comprising a nucleotide sequence represented by anyof SEQ ID NOs: 181 to 194,(m) a polynucleotide consisting of a nucleotide sequence complementaryto a nucleotide sequence represented by any of SEQ ID NOs: 181 to 194 ora nucleotide sequence derived from the nucleotide sequence by thereplacement of u with t, a variant thereof, a derivative thereof, or afragment thereof comprising 15 or more consecutive nucleotides,(n) a polynucleotide comprising a nucleotide sequence complementary to anucleotide sequence represented by any of SEQ ID NOs: 181 to 194 or anucleotide sequence derived from the nucleotide sequence by thereplacement of u with t, and(o) a polynucleotide hybridizing under stringent conditions to any ofthe polynucleotides (k) to (n).

(10) The kit according to any one of (1) to (9), wherein the kitcomprises at least two or more nucleic acids capable of specificallybinding to at least two or more polynucleotides, respectively, selectedfrom the group consisting of all of the colorectal cancer markersaccording to (1) or (2).

(11) A device for the detection of colorectal cancer, comprising anucleic acid capable of specifically binding to at least one or morepolynucleotide(s) selected from the group consisting of colorectalcancer markers miR-6726-5p, miR-4257, miR-6787-5p, miR-6780b-5p,miR-3131, miR-7108-5p, miR-1343-3p, miR-1247-3p, miR-4651, miR-6757-5p,miR-3679-5p, miR-7641, miR-6746-5p, miR-8072, miR-6741-5p, miR-1908-5p,miR-6857-5p, miR-4746-3p, miR-744-5p, miR-4792, miR-564, miR-6791-5p,miR-6825-5p, miR-6826-5p, miR-4665-3p, miR-4467, miR-3188, miR-6125,miR-6756-5p, miR-1228-3p, miR-8063, miR-8069, miR-6875-5p, miR-3185,miR-4433b-3p, miR-6887-5p, miR-128-1-5p, miR-6724-5p, miR-1914-3p,miR-1225-5p, miR-4419b, miR-7110-5p, miR-187-5p, miR-3184-5p,miR-204-3p, miR-5572, miR-6729-5p, miR-615-5p, miR-6749-5p, miR-6515-3p,miR-3937, miR-6840-3p, miR-6893-5p, miR-4728-5p, miR-6717-5p,miR-7113-3p, miR-4665-5p, miR-642b-3p, miR-7109-5p, miR-6842-5p,miR-4442, miR-4433-3p, miR-4707-5p, miR-6126, miR-4449, miR-4706,miR-1913, miR-602, miR-939-5p, miR-4695-5p, miR-711, miR-6816-5p,miR-4632-5p, miR-6721-5p, miR-7847-3p, miR-6132, miR-887-3p,miR-3679-3p, miR-6784-5p, miR-1249, miR-937-5p, miR-5195-3p,miR-6732-5p, miR-4417, miR-4281, miR-4734, miR-6766-3p, miR-663a,miR-4513, miR-6781-5p, miR-1227-5p, miR-6845-5p, miR-6798-5p,miR-3620-5p, miR-1915-5p, miR-4294, miR-642a-3p, miR-371a-5p, miR-940,miR-4450, miR-4723-5p, miR-1469, miR-6861-5p, miR-7975, miR-6879-5p,miR-6802-5p, miR-1268b, miR-663b, miR-125a-3p, miR-2861, miR-6088,miR-4758-5p, miR-296-3p, miR-6738-5p, miR-671-5p, miR-4454, miR-4516,miR-7845-5p, miR-4741, miR-92b-5p, miR-6795-5p, miR-6805-3p,miR-4725-3p, miR-6782-5p, miR-4688, miR-6850-5p, miR-6777-5p,miR-6785-5p, miR-7106-5p, miR-3663-3p, miR-6131, miR-1915-3p, miR-4532,miR-6820-5p, miR-4689, miR-4638-5p, miR-3656, miR-3621, miR-6769b-5p,miR-149-3p, miR-23b-3p, miR-3135b, miR-6848-5p, miR-6769a-5p, miR-4327,miR-6765-3p, miR-6716-5p, miR-6877-5p, miR-6727-5p, miR-4534, miR-614,miR-1202, miR-575, miR-6870-5p, miR-6722-3p, miR-7977, miR-4649-5p,miR-4675, miR-6075, miR-6779-5p, miR-4271, miR-3196, miR-6803-5p,miR-6789-5p, miR-4648, miR-4508, miR-4749-5p, miR-4505, miR-5698,miR-1199-5p, miR-4763-3p, miR-6836-3p, miR-3195, miR-718, miR-3178,miR-638, miR-4497, miR-6085, miR-6752-5p and miR-135a-3p.

(12) The device according to (11), wherein miR-6726-5p ishsa-miR-6726-5p, miR-4257 is hsa-miR-4257, miR-6787-5p ishsa-miR-6787-5p, miR-6780b-5p is hsa-miR-6780b-5p, miR-3131 ishsa-miR-3131, miR-7108-5p is hsa-miR-7108-5p, miR-1343-3p ishsa-miR-1343-3p, miR-1247-3p is hsa-miR-1247-3p, miR-4651 ishsa-miR-4651, miR-6757-5p is hsa-miR-6757-5p, miR-3679-5p ishsa-miR-3679-5p, miR-7641 is hsa-miR-7641, miR-6746-5p ishsa-miR-6746-5p, miR-8072 is hsa-miR-8072, miR-6741-5p ishsa-miR-6741-5p, miR-1908-5p is hsa-miR-1908-5p, miR-6857-5p ishsa-miR-6857-5p, miR-4746-3p is hsa-miR-4746-3p, miR 5p ishsa-miR-744-5p, miR-4792 is hsa-miR-4792, miR-564 is hsa-miR-564,miR-6791-5p is hsa-miR-6791-5p, miR-6825-5p is hsa-miR-6825-5p,miR-6826-5p is hsa-miR-6826-5p, miR-4665-3p is hsa-miR-4665-3p, miR-4467is hsa-miR-4467, miR-3188 is hsa-miR-3188, miR-6125 is hsa-miR-6125,miR-6756-5p is hsa-miR-6756-5p, miR-1228-3p is hsa-miR-1228-3p, miR-8063is hsa-miR-8063, miR-8069 is hsa-miR-8069, miR-6875-5p ishsa-miR-6875-5p, miR-3185 is hsa-miR-3185, miR-4433b-3p ishsa-miR-4433b-3p, miR-6887-5p is hsa-miR-6887-5p, miR-128-1-5p ishsa-miR-128-1-5p, miR-6724-5p is hsa-miR-6724-5p, miR-1914-3p ishsa-miR-1914-3p, miR-1225-5p is hsa-miR-1225-5p, miR-4419b ishsa-miR-4419b, miR-7110-5p is hsa-miR-7110-5p, miR-187-5p ishsa-miR-187-5p, miR-3184-5p is hsa-miR-3184-5p, miR-204-3p ishsa-miR-204-3p, miR-5572 is hsa-miR-5572, miR-6729-5p ishsa-miR-6729-5p, miR-615-5p is hsa-miR-615-5p, miR-6749-5p ishsa-miR-6749-5p, miR-6515-3p is hsa-miR-6515-3p, miR-3937 ishsa-miR-3937, miR-6840-3p is hsa-miR-6840-3p, miR-6893-5p ishsa-miR-6893-5p, miR-4728-5p is hsa-miR-4728-5p, miR-6717-5p ishsa-miR-6717-5p, miR-7113-3p is hsa-miR-7113-3p, miR-4665-5p ishsa-miR-4665-5p, miR-642b-3p is hsa-miR-642b-3p, miR-7109-5p ishsa-miR-7109-5p, miR-6842-5p is hsa-miR-6842-5p, miR-4442 ishsa-miR-4442, miR-4433-3p is hsa-miR-4433-3p, miR-4707-5p ishsa-miR-4707-5p, miR-6126 is hsa-miR-6126, miR-4449 is hsa-miR-4449,miR-4706 is hsa-miR-4706, miR-1913 is hsa-miR-1913, miR-602 ishsa-miR-602, miR-939-5p is hsa-miR-939-5p, miR-4695-5p ishsa-miR-4695-5p, miR-711 is hsa-miR-711, miR-6816-5p is hsa-miR-6816-5p,miR-4632-5p is hsa-miR-4632-5p, miR-6721-5p is hsa-miR-6721-5p,miR-7847-3p is hsa-miR-7847-3p, miR-6132 is hsa-miR-6132, miR-887-3p ishsa-miR-887-3p, miR-3679-3p is hsa-miR-3679-3p, miR-6784-5p ishsa-miR-6784-5p, miR-1249 is hsa-miR-1249, miR-937-5p is hsa-miR-937-5p,miR-5195-3p is hsa-miR-5195-3p, miR-6732-5p is hsa-miR-6732-5p, miR-4417is hsa-miR-4417, miR-4281 is hsa-miR-4281, miR-4734 is hsa-miR-4734,miR-6766-3p is hsa-miR-6766-3p, miR-663a is hsa-miR-663a, miR-4513 ishsa-miR-4513, miR-6781-5p is hsa-miR-6781-5p, miR-1227-5p ishsa-miR-1227-5p, miR-6845-5p is hsa-miR-6845-5p, miR-6798-5p ishsa-miR-6798-5p, miR-3620-5p is hsa-miR-3620-5p, miR-1915-5p ishsa-miR-1915-5p, miR-4294 is hsa-miR-4294, miR-642a-3p ishsa-miR-642a-3p, miR-371a-5p is hsa-miR-371a-5p, miR-940 is hsa-miR-940,miR-4450 is hsa-miR-4450, miR-4723-5p is hsa-miR-4723-5p, miR-1469 ishsa-miR-1469, miR-6861-5p is hsa-miR-6861-5p, miR-7975 is hsa-miR-7975,miR-6879-5p is hsa-miR-6879-5p, miR-6802-5p is hsa-miR-6802-5p,miR-1268b is hsa-miR-1268b, miR-663b is hsa-miR-663b, miR-125a-3p ishsa-miR-125a-3p, miR-2861 is hsa-miR-2861, miR-6088 is hsa-miR-6088,miR-4758-5p is hsa-miR-4758-5p, miR-296-3p is hsa-miR-296-3p,miR-6738-5p is hsa-miR-6738-5p, miR-671-5p is hsa-miR-671-5p, miR-4454is hsa-miR-4454, miR-4516 is hsa-miR-4516, miR-7845-5p ishsa-miR-7845-5p, miR-4741 is hsa-miR-4741, miR-92b-5p is hsa-miR-92b-5p,miR-6795-5p is hsa-miR-6795-5p, miR-6805-3p is hsa-miR-6805-3p,miR-4725-3p is hsa-miR-4725-3p, miR-6782-5p is hsa-miR-6782-5p, miR-4688is hsa-miR-4688, miR-6850-5p is hsa-miR-6850-5p, miR-6777-5p ishsa-miR-6777-5p, miR-6785-5p is hsa-miR-6785-5p, miR-7106-5p ishsa-miR-7106-5p, miR-3663-3p is hsa-miR-3663-3p, miR-6131 ishsa-miR-6131, miR-1915-3p is hsa-miR-1915-3p, miR-4532 is hsa-miR-4532,miR-6820-5p is hsa-miR-6820-5p, miR-4689 is hsa-miR-4689, miR-4638-5p ishsa-miR-4638-5p, miR-3656 is hsa-miR-3656, miR-3621 is hsa-miR-3621,miR-6769b-5p is hsa-miR-6769b-5p, miR-149-3p is hsa-miR-149-3p,miR-23b-3p is hsa-miR-23b-3p, miR-3135b is hsa-miR-3135b, miR-6848-5p ishsa-miR-6848-5p, miR-6769a-5p is hsa-miR-6769a-5p, miR-4327 ishsa-miR-4327, miR-6765-3p is hsa-miR-6765-3p, miR-6716-5p ishsa-miR-6716-5p, miR-6877-5p is hsa-miR-6877-5p, miR-6727-5p ishsa-miR-6727-5p, miR-4534 is hsa-miR-4534, miR-614 is hsa-miR-614,miR-1202 is hsa-miR-1202, miR-575 is hsa-miR-575, miR-6870-5p ishsa-miR-6870-5p, miR-6722-3p is hsa-miR-6722-3p, miR-7977 ishsa-miR-7977, miR-4649-5p is hsa-miR-4649-5p, miR-4675 is hsa-miR-4675,miR-6075 is hsa-miR-6075, miR-6779-5p is hsa-miR-6779-5p, miR-4271 ishsa-miR-4271, miR-3196 is hsa-miR-3196, miR-6803-5p is hsa-miR-6803-5p,miR-6789-5p is hsa-miR-6789-5p, miR-4648 is hsa-miR-4648, miR-4508 ishsa-miR-4508, miR-4749-5p is hsa-miR-4749-5p, miR-4505 is hsa-miR-4505,miR-5698 is hsa-miR-5698, miR-1199-5p is hsa-miR-1199-5p, miR-4763-3p ishsa-miR-4763-3p, miR-6836-3p is hsa-miR-6836-3p, miR-3195 ishsa-miR-3195, miR-718 is hsa-miR-718, miR-3178 is hsa-miR-3178, miR-638is hsa-miR-638, miR-4497 is hsa-miR-4497, miR-6085 is hsa-miR-6085,miR-6752-5p is hsa-miR-6752-5p, and miR-135a-3p is hsa-miR-135a-3p.

(13) The device according to (11) or (12), wherein the nucleic acid is apolynucleotide selected from the group consisting of the followingpolynucleotides (a) to (e):

(a) a polynucleotide consisting of a nucleotide sequence represented byany of SEQ ID NOs: 1 to 171 and 606 to 614 or a nucleotide sequencederived from the nucleotide sequence by the replacement of u with t, avariant thereof, a derivative thereof, or a fragment thereof comprising15 or more consecutive nucleotides,(b) a polynucleotide comprising a nucleotide sequence represented by anyof SEQ ID NOs: 1 to 171 and 606 to 614,(c) a polynucleotide consisting of a nucleotide sequence complementaryto a nucleotide sequence represented by any of SEQ ID NOs: 1 to 171 and606 to 614 or a nucleotide sequence derived from the nucleotide sequenceby the replacement of u with t, a variant thereof, a derivative thereof,or a fragment thereof comprising 15 or more consecutive nucleotides,(d) a polynucleotide comprising a nucleotide sequence complementary to anucleotide sequence represented by any of SEQ ID NOs: 1 to 171 and 606to 614 or a nucleotide sequence derived from the nucleotide sequence bythe replacement of u with t, and(e) a polynucleotide hybridizing under stringent conditions to any ofthe polynucleotides (a) to (d).

(14) The device according to any of (11) to (13), wherein the devicefurther comprises a nucleic acid capable of specifically binding to atleast one or more polynucleotide(s) selected from the group consistingof other colorectal cancer markers miR-1231, miR-1233-5p, miR-150-3p,miR-1225-3p, miR-92a-2-5p, miR-423-5p, miR-1268a, miR-128-2-5p andmiR-24-3p.

(15) The device according to (14), wherein miR-1231 is hsa-miR-1231,miR-1233-5p is hsa-miR-1233-5p, miR-150-3p is hsa-miR-150-3p,miR-1225-3p is hsa-miR-1225-3p, miR-92a-2-5p is hsa-miR-92a-2-5p,miR-423-5p is hsa-miR-423-5p, miR-1268a is hsa-miR-1268a, miR-128-2-5pis hsa-miR-128-2-5p, and miR-24-3p is hsa-miR-24-3p.

(16) The device according to (14) or (15), wherein the nucleic acid is apolynucleotide selected from the group consisting of the followingpolynucleotides (f) to (j):

(f) a polynucleotide consisting of a nucleotide sequence represented byany of SEQ ID NOs: 172 to 180 or a nucleotide sequence derived from thenucleotide sequence by the replacement of u with t, a variant thereof, aderivative thereof, or a fragment thereof comprising 15 or moreconsecutive nucleotides,(g) a polynucleotide comprising a nucleotide sequence represented by anyof SEQ ID NOs: 172 to 180,(h) a polynucleotide consisting of a nucleotide sequence complementaryto a nucleotide sequence represented by any of SEQ ID NOs: 172 to 180 ora nucleotide sequence derived from the nucleotide sequence by thereplacement of u with t, a variant thereof, a derivative thereof, or afragment thereof comprising 15 or more consecutive nucleotides,(i) a polynucleotide comprising a nucleotide sequence complementary to anucleotide sequence represented by any of SEQ ID NOs: 172 to 180 or anucleotide sequence derived from the nucleotide sequence by thereplacement of u with t, and(j) a polynucleotide hybridizing under stringent conditions to any ofthe polynucleotides (f) to (i).

(17) The device according to any of (11) to (16), wherein the devicefurther comprises a nucleic acid capable of specifically binding to atleast one or more polynucleotide(s) selected from the group consistingof other colorectal cancer markers miR-4697-5p, miR-3197, miR-675-5p,miR-4486, miR-7107-5p, miR-23a-3p, miR-4667-5p, miR-451a, miR-3940-5p,miR-8059, miR-6813-5p, miR-4492, miR-4476, and miR-6090.

(18) The device according to (17), wherein miR-4697-5p ishsa-miR-4697-5p, miR-3197 is hsa-miR-3197, miR-675-5p is hsa-miR-675-5p,miR-4486 is hsa-miR-4486, miR-7107-5p is hsa-miR-7107-5p, miR-23a-3p ishsa-miR-23a-3p, miR-4667-5p is hsa-miR-4667-5p, miR-451a ishsa-miR-451a, miR-3940-5p is hsa-miR-3940-5p, miR-8059 is hsa-miR-8059,miR-6813-5p is hsa-miR-6813-5p, miR-4492 is hsa-miR-4492, miR-4476 ishsa-miR-4476, and miR-6090 is hsa-miR-6090.

(19) The device according to (17) or (18), wherein the nucleic acid is apolynucleotide selected from the group consisting of the followingpolynucleotides (k) to (o):

(k) a polynucleotide consisting of a nucleotide sequence represented byany of SEQ ID NOs: 181 to 194 or a nucleotide sequence derived from thenucleotide sequence by the replacement of u with t, a variant thereof, aderivative thereof, or a fragment thereof comprising 15 or moreconsecutive nucleotides,(l) a polynucleotide comprising a nucleotide sequence represented by anyof SEQ ID NOs: 181 to 194,(m) a polynucleotide consisting of a nucleotide sequence complementaryto a nucleotide sequence represented by any of SEQ ID NOs: 181 to 194 ora nucleotide sequence derived from the nucleotide sequence by thereplacement of u with t, a variant thereof, a derivative thereof, or afragment thereof comprising 15 or more consecutive nucleotides,(n) a polynucleotide comprising a nucleotide sequence complementary to anucleotide sequence represented by any of SEQ ID NOs: 181 to 194 or anucleotide sequence derived from the nucleotide sequence by thereplacement of u with t, and(o) a polynucleotide hybridizing under stringent conditions to any ofthe polynucleotides (k) to (n).

(20) The device according to any one of (11) to (19), wherein the deviceis a device for measurement by a hybridization technique.

(21) The device according to (20), wherein the hybridization techniqueis a nucleic acid array technique.

(22) The device according to any one of (11) to (21), wherein the devicecomprises at least two or more nucleic acids capable of specificallybinding to at least two or more polynucleotides, respectively, selectedfrom all of the colorectal cancer markers according to (11) or (12).

(23) A method for detecting colorectal cancer, comprising measuring anexpression level of a target nucleic acid in a sample from a subjectusing a kit according to any one of (1) to (10) or a device according toany one of (11) to (22), and evaluating in vitro whether or not thesubject has colorectal cancer using both of the measured expressionlevel and a control expression level in a sample from a healthy subjectmeasured in the same way.

(24) The method according to (23), wherein the subject is a human.

(25) The method according to (23) or (24), wherein the sample is blood,serum, or plasma.

Definition of Term

The terms used herein are defined as follows.

Abbreviations or terms such as nucleotide, polynucleotide, DNA, and RNAabide by “Guidelines for the preparation of specification which containnucleotide and/or amino acid sequences” (edited by Japan Patent Office)and common use in the art.

The term “polynucleotide” used herein is used for a nucleic acidincluding any of RNA, DNA, and RNA/DNA (chimera). The DNA includes anyof cDNA, genomic DNA, and synthetic DNA. The RNA includes any of totalRNA, mRNA, rRNA, miRNA, siRNA, snoRNA, snRNA, non-coding RNA andsynthetic RNA. Here, the “synthetic DNA” and the “synthetic RNA” referto DNA and RNA artificially prepared using, for example, an automaticnucleic acid synthesizer, on the basis of predetermined nucleotidesequences (which may be any of natural and non-natural sequences). The“non-natural sequence” is intended to be used in a broad sense andincludes, for example, a sequence comprising substitution, deletion,insertion, and/or addition of one or more nucleotide(s) (i.e., a variantsequence) and a sequence comprising one or more modified nucleotide(s)(i.e., a modified sequence), which are different from the naturalsequence. Here, the term “polynucleotide” is used interchangeably withthe term “nucleic acid”.

The term “fragment” used herein is a polynucleotide having a nucleotidesequence that consists of a consecutive portion of a polynucleotide anddesirably has a length of 15 or more nucleotides, preferably 17 or morenucleotides, more preferably 19 or more nucleotides.

The term “gene” used herein is intended to include not only RNA anddouble-stranded DNA but each single-stranded DNA such as a plus strand(or a sense strand) or a complementary strand (or an antisense strand)that constitutes a duplex. The gene is not particularly limited by itslength.

Thus, the “gene” used herein includes any of double-stranded DNAincluding human genomic DNA, single-stranded DNA (plus strand),single-stranded DNA having a sequence complementary to the plus strand(complementary strand) including cDNA, microRNA (miRNA), and theirfragments, and transcripts, unless otherwise specified. The “gene”includes not only a “gene” represented by a particular nucleotidesequence (or SEQ ID NO) but “nucleic acids” encoding RNAs havingbiological functions equivalent to RNA encoded by the gene, for example,a congener (i.e., a homolog or an ortholog), a variant (e.g., a geneticpolymorph), and a derivative. Specific examples of such a “nucleic acid”encoding a congener, a variant, or a derivative can include a “nucleicacid” having a nucleotide sequence hybridizing under stringentconditions described later to a complementary sequence of a nucleotidesequence represented by any of SEQ ID NOs: 1 to 635 or a nucleotidesequence derived from the nucleotide sequence by the replacement of uwith t. The “gene” is not particularly limited by its functional regionand can contain, for example, an expression control region, a codingregion, an exon, or an intron. The “gene” may be contained in a cell ormay exist alone after being released into the outside of a cell.Alternatively, the “gene” may be in a state enclosed in a vesicle calledexosome.

The term “exosome” used herein is a vesicle that is encapsulated by alipid bilayer and secreted from a cell. The exosome is derived from amultivesicular endosome and may incorporate a biomaterial such as a“gene” (e.g., RNA or DNA) or a protein when released into anextracellular environment. The exosome is known to be contained in abody fluid such as blood, serum, plasma, or lymph.

The term “transcript” used herein refers to RNA synthesized with the DNAsequence of a gene as a template. RNA polymerase binds to a site calledpromoter which is located upstream of the gene and adds ribonucleotidescomplementary to the nucleotide sequence of the DNA to the 3′ end tosynthesize RNA. This RNA contains not only the gene itself but the wholesequence from a transcription initiation site to the end of a polyAsequence, including an expression regulatory region, a coding region, anexon, or an intron.

The term “microRNA (miRNA)” used herein is intended to mean a 15- to25-nucleotide non-coding RNA that is transcribed as an RNA precursorhaving a hairpin-like structure, cleaved by a dsRNA-cleaving enzymehaving RNase III cleavage activity, and integrated into a proteincomplex called RISC, and is involved in the suppression of translationof mRNA, unless otherwise specified. The term “miRNA” used hereinincludes not only a “miRNA” represented by a particular nucleotidesequence (or SEQ ID NO) but a precursor of the “miRNA” (pre-miRNA orpri-miRNA), and miRNAs having biological functions equivalent thereto,for example, a congener (i.e., a homolog or an ortholog), a variant(e.g., a genetic polymorph), and a derivative. Such a precursor, acongener, a variant, or a derivative can be specifically identifiedusing miRBase Release 20 (http://www.mirbase.org/), and examples thereofcan include a “miRNA” having a nucleotide sequence hybridizing understringent conditions described later to a complementary sequence of anyparticular nucleotide sequence represented by any of SEQ ID NOs: 1 to635. The term “miRNA” used herein may be a gene product of a miR gene.Such a gene product includes a mature miRNA (e.g., a 15- to25-nucleotide or 19- to 25-nucleotide non-coding RNA involved in thesuppression of translation of mRNA as described above) or a miRNAprecursor (e.g., pre-miRNA or pri-miRNA as described above).

The term “probe” used herein includes a polynucleotide that is used forspecifically detecting RNA resulting from the expression of a gene or apolynucleotide derived from the RNA, and/or a polynucleotidecomplementary thereto.

The term “primer” used herein includes a polynucleotide thatspecifically recognizes and amplifies RNA resulting from the expressionof a gene or a polynucleotide derived from the RNA, and/or apolynucleotide complementary thereto.

In this context, the complementary polynucleotide (complementary strandor reverse strand) means a polynucleotide in a complementary baserelationship of A:T (U) and G:C base pairs with the full-length sequenceof a polynucleotide consisting of a nucleotide sequence defined by anyof SEQ ID NOs: 1 to 635 or a nucleotide sequence derived from thenucleotide sequence by the replacement of u with t, or a partialsequence thereof (here, this full-length or partial sequence is referredto as a plus strand for the sake of convenience). However, such acomplementary strand is not limited to a sequence completelycomplementary to the nucleotide sequence of the target plus strand andmay have a complementary relationship to an extent that permitshybridization under stringent conditions to the target plus strand.

The term “stringent conditions” used herein refers to conditions underwhich a nucleic acid probe hybridizes to its target sequence to a largerextent (e.g., a measurement value equal to or larger than a mean ofbackground measurement values+a standard deviation of the backgroundmeasurement values×2) than that for other sequences. The stringentconditions are dependent on a sequence and differ depending on anenvironment where hybridization is performed. A target sequencecomplementary 100% to the nucleic acid probe can be identified bycontrolling the stringency of hybridization and/or washing conditions.Specific examples of the “stringent conditions” will be mentioned later.

The term “Tm value” used herein means a temperature at which thedouble-stranded moiety of a polynucleotide is denatured into singlestrands so that the double strands and the single strands exist at aratio of 1:1.

The term “variant” used herein means, in the case of a nucleic acid, anatural variant attributed to polymorphism, mutation, or the like; avariant that contains the deletion, substitution, addition, or insertionof 1 or 2 or more nucleotides in a nucleotide sequence represented byany of SEQ ID NOs: 1 to 194 and 606 to 614 or a nucleotide sequencederived from the nucleotide sequence by the replacement of u with t, ora partial sequence thereof; a variant that exhibits % identity ofapproximately 90% or higher, approximately 95% or higher, approximately97% or higher, approximately 98% or higher, approximately 99% or higherto each of these nucleotide sequences or the partial sequence thereof;or a nucleic acid hybridizing under the stringent conditions definedabove to a polynucleotide or an oligonucleotide comprising each of thesenucleotide sequences or the partial sequence thereof.

The term “several” used herein means an integer of approximately 10, 9,8, 7, 6, 5, 4, 3, or 2.

The variant used herein can be prepared by use of a well-known techniquesuch as site-directed mutagenesis or PCR-based mutagenesis.

The term “percent (%) identity” used herein can be determined with orwithout an introduced gap, using a protein or gene search system basedon BLAST or FASTA described above (Zheng Zhang et al., 2000, J. Comput.Biol., Vol. 7, p. 203-214; Altschul, S. F. et al., 1990, Journal ofMolecular Biology, Vol. 215, p. 403-410; and Pearson, W. R. et al.,1988, Proc. Natl. Acad. Sci. U.S.A. Vol. 85, p. 2444-2448).

The term “derivative” used herein is meant to include a modified nucleicacid, for example, a derivative that is labeled with a fluorophore orthe like, a derivative containing a modified nucleotide (e.g., anucleotide containing a group such as halogen, alkyl such as methyl,alkoxy such as methoxy, thio, or carboxymethyl, and a nucleotide thathas undergone base rearrangement, double bond saturation, deamination,replacement of an oxygen molecule with a sulfur atom, etc.), PNA(peptide nucleic acid; Nielsen, P. E. et al., 1991, Science, Vol. 254,p. 1497-500), and LNA (locked nucleic acid; Obika, S. et al., 1998,Tetrahedron Lett., Vol. 39, p. 5401-5404) without any limitation.

As used herein, the “nucleic acid” capable of specifically binding to apolynucleotide selected from the colorectal cancer marker miRNAsdescribed above is a synthesized or prepared nucleic acid andspecifically includes a “nucleic acid probe” or a “primer”. The “nucleicacid” is utilized directly or indirectly for detecting the presence orabsence of colorectal cancer in a subject, for diagnosing the presenceor absence of colorectal cancer, for diagnosing the severity ofcolorectal cancer, the presence or absence of amelioration or the degreeof amelioration of colorectal cancer, or the sensitivity to treatmentfor colorectal cancer, or for screening for a candidate substance usefulin the prevention, amelioration, or treatment of colorectal cancer. The“nucleic acid” includes a nucleotide, an oligonucleotide, and apolynucleotide capable of specifically recognizing and binding to atranscript represented by any of SEQ ID NOs: 1 to 635 or a syntheticcDNA nucleic acid thereof in vivo, particularly, in a sample such as abody fluid (e.g., blood or urine), in relation to the development ofcolorectal cancer. The nucleotide, the oligonucleotide, and thepolynucleotide can be effectively used as probes for detecting theaforementioned gene expressed in vivo, in tissues, in cells, or the likeon the basis of the properties described above, or as primers foramplifying the aforementioned gene expressed in vivo.

The term “capable of specifically binding” used herein means that thenucleic acid probe or the primer used in the present invention binds toa particular target nucleic acid and cannot substantially bind to othernucleic acids.

The term “detection” used herein is interchangeable with the term“examination”, “measurement”, “detection”, or “decision support”. Asused herein, the term “evaluation” is meant to include diagnosing orevaluation-supporting on the basis of examination results or measurementresults.

The term “subject” used herein means a mammal such as a primateincluding a human and a chimpanzee, a pet animal including a dog and acat, a livestock animal including cattle, a horse, sheep, and a goat,and a rodent including a mouse and a rat. The term “healthy subject”also means such a mammal without the cancer to be detected.

The term “P” or “P value” used herein refers to a probability at which amore extreme statistic than that is actually calculated from data undernull hypothesis is observed in a statistical test. Thus, smaller “P” or“P value” means more significant difference between subjects to becompared.

The term “sensitivity” used herein means a value of (the number of truepositives)/(the number of true positives+the number of false negatives).High sensitivity allows colorectal cancer to be detected early, leadingto the complete resection of cancer sites and reduction in the rate ofrecurrence.

The term “specificity” used herein means a value of (the number of truenegatives)/(the number of true negatives+the number of false positives).High specificity prevents needless extra examination for healthysubjects misjudged as being colorectal cancer patients, leading toreduction in burden on patients and reduction in medical expense.

The term “accuracy” used herein means a value of (the number of truepositives+the number of true negatives)/(the total number of cases). Theaccuracy indicates the ratio of samples that correctly identified indiscriminant results to all samples, and serves as a primary index forevaluating detection performance.

As used herein, the “sample” that is subject to determination,detection, or diagnosis refers to a tissue and a biological material inwhich the expression of the gene of the present invention varies ascolorectal cancer develops, colorectal cancer progresses, andtherapeutic effects on colorectal cancer are exerted. Specifically, the“sample” refers to a large intestine tissue, a vascular channel aroundthe large intestine, lymph node, and organ, an organ suspected of havingmetastasis, the skin, a body fluid such as blood, urine, saliva, sweat,or tissue exudates, serum or plasma prepared from blood, feces, hair,and the like. The “sample” further refers to a biological sampleextracted therefrom, specifically, a gene such as RNA or miRNA.

The term “hsa-miR-6726-5p gene” or “hsa-miR-6726-5p” used hereinincludes the hsa-miR-6726-5p gene (miRBase Accession No. MIMAT0027353)described in SEQ ID NO: 1, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6726-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6726” (miRBase Accession No. MI0022571, SEQ IDNO: 195) having a hairpin-like structure is known as a precursor of“hsa-miR-6726-5p”.

The term “hsa-miR-4257 gene” or “hsa-miR-4257” used herein includes thehsa-miR-4257 gene (miRBase Accession No. MIMAT0016878) described in SEQID NO: 2, a homolog or an ortholog of a different organism species, andthe like. The hsa-miR-4257 gene can be obtained by a method described inGoff L A et al., 2009, PLoS One, Vol. 4, e7192. Also, “hsa-mir-4257”(miRBase Accession No. MI0015856, SEQ ID NO: 196) having a hairpin-likestructure is known as a precursor of “hsa-miR-4257”.

The term “hsa-miR-6787-5p gene” or “hsa-miR-6787-5p” used hereinincludes the hsa-miR-6787-5p gene (miRBase Accession No. MIMAT0027474)described in SEQ ID NO: 3, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6787-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6787” (miRBase Accession No. MI0022632, SEQ IDNO: 197) having a hairpin-like structure is known as a precursor of“hsa-miR-6787-5p”.

The term “hsa-miR-6780b-5p gene” or “hsa-miR-6780b-5p” used hereinincludes the hsa-miR-6780b-5p gene (miRBase Accession No. MIMAT0027572)described in SEQ ID NO: 4, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6780b-5p gene can beobtained by a method described in Ladewig E et al., 2012, Genome Res,Vol. 22, p. 1634-1645. Also, “hsa-mir-6780b” (miRBase Accession No.MI0022681, SEQ ID NO: 198) having a hairpin-like structure is known as aprecursor of “hsa-miR-6780b-5p”.

The term “hsa-miR-3131 gene” or “hsa-miR-3131” used herein includes thehsa-miR-3131 gene (miRBase Accession No. MIMAT0014996) described in SEQID NO: 5, a homolog or an ortholog of a different organism species, andthe like. The hsa-miR-3131 gene can be obtained by a method described inStark M S et al., 2010, PLoS One, Vol. 5, e9685. Also, “hsa-mir-3131”(miRBase Accession No. MI0014151, SEQ ID NO: 199) having a hairpin-likestructure is known as a precursor of “hsa-miR-3131”.

The term “hsa-miR-7108-5p gene” or “hsa-miR-7108-5p” used hereinincludes the hsa-miR-7108-5p gene (miRBase Accession No. MIMAT0028113)described in SEQ ID NO: 6, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-7108-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-7108” (miRBase Accession No. MI0022959, SEQ IDNO: 200) having a hairpin-like structure is known as a precursor of“hsa-miR-7108-5p”.

The term “hsa-miR-1343-3p gene” or “hsa-miR-1343-3p” used hereinincludes the hsa-miR-1343-3p gene (miRBase Accession No. MIMAT0019776)described in SEQ ID NO: 7, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-1343-3p gene can be obtainedby a method described in Persson H et al., 2011, Cancer Res, Vol. 71, p.78-86. Also, “hsa-mir-1343” (miRBase Accession No. MI0017320, SEQ ID NO:201) having a hairpin-like structure is known as a precursor of“hsa-miR-1343-3p”.

The term “hsa-miR-1247-3p gene” or “hsa-miR-1247-3p” used hereinincludes the hsa-miR-1247-3p gene (miRBase Accession No. MIMAT0022721)described in SEQ ID NO: 8, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-1247-3p gene can be obtainedby a method described in Morin R D et al., 2008, Genome Res, Vol. 18, p.610-621. Also, “hsa-mir-1247” (miRBase Accession No. MI0006382, SEQ IDNO: 202) having a hairpin-like structure is known as a precursor of“hsa-miR-1247-3p”.

The term “hsa-miR-4651 gene” or “hsa-miR-4651” used herein includes thehsa-miR-4651 gene (miRBase Accession No. MIMAT0019715) described in SEQID NO: 9, a homolog or an ortholog of a different organism species, andthe like. The hsa-miR-4651 gene can be obtained by a method described inPersson H et al., 2011, Cancer Res, Vol. 71, p. 78-86. Also,“hsa-mir-4651” (miRBase Accession No. MI0017279, SEQ ID NO: 203) havinga hairpin-like structure is known as a precursor of “hsa-miR-4651”.

The term “hsa-miR-6757-5p gene” or “hsa-miR-6757-5p” used hereinincludes the hsa-miR-6757-5p gene (miRBase Accession No. MIMAT0027414)described in SEQ ID NO: 10, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6757-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6757” (miRBase Accession No. MI0022602, SEQ IDNO: 204) having a hairpin-like structure is known as a precursor of“hsa-miR-6757-5p”.

The term “hsa-miR-3679-5p gene” or “hsa-miR-3679-5p” used hereinincludes the hsa-miR-3679-5p gene (miRBase Accession No. MIMAT0018104)described in SEQ ID NO: 11, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-3679-5p gene can be obtainedby a method described in Creighton C J et al., 2010, PLoS One, Vol. 5,e9637. Also, “hsa-mir-3679” (miRBase Accession No. MI0016080, SEQ ID NO:205) having a hairpin-like structure is known as a precursor of“hsa-miR-3679-5p”.

The term “hsa-miR-7641 gene” or “hsa-miR-7641” used herein includes thehsa-miR-7641 gene (miRBase Accession No. MIMAT0029782) described in SEQID NO: 12, a homolog or an ortholog of a different organism species, andthe like. The hsa-miR-7641 gene can be obtained by a method described inYoo J K et al., 2013, Arch Pharm Res, Vol. 36, p. 353-358. Also,“hsa-mir-7641-1” and “hsa-mir-7641-2” (miRBase Accession Nos. MI0024975and MI0024976, SEQ ID NOs: 206 and 207) having a hairpin-like structureare known as precursors of “hsa-miR-7641”.

The term “hsa-miR-6746-5p gene” or “hsa-miR-6746-5p” used hereinincludes the hsa-miR-6746-5p gene (miRBase Accession No. MIMAT0027392)described in SEQ ID NO: 13, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6746-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6746” (miRBase Accession No. MI0022591, SEQ IDNO: 208) having a hairpin-like structure is known as a precursor of“hsa-miR-6746-5p”.

The term “hsa-miR-8072 gene” or “hsa-miR-8072” used herein includes thehsa-miR-8072 gene (miRBase Accession No. MIMAT0030999) described in SEQID NO: 14, a homolog or an ortholog of a different organism species, andthe like. The hsa-miR-8072 gene can be obtained by a method described inWang H J et al., 2013, Shock, Vol. 39, p. 480-487. Also, “hsa-mir-8072”(miRBase Accession No. MI0025908, SEQ ID NO: 209) having a hairpin-likestructure is known as a precursor of “hsa-miR-8072”.

The term “hsa-miR-6741-5p gene” or “hsa-miR-6741-5p” used hereinincludes the hsa-miR-6741-5p gene (miRBase Accession No. MIMAT0027383)described in SEQ ID NO: 15, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6741-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6741” (miRBase Accession No. MI0022586, SEQ IDNO: 210) having a hairpin-like structure is known as a precursor of“hsa-miR-6741-5p”.

The term “hsa-miR-1908-5p gene” or “hsa-miR-1908-5p” used hereinincludes the hsa-miR-1908-5p gene (miRBase Accession No. MIMAT0007881)described in SEQ ID NO: 16, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-1908-5p gene can be obtainedby a method described in Bar M et al., 2008, Stem Cells, Vol. 26, p.2496-2505. Also, “hsa-mir-1908” (miRBase Accession No. MI0008329, SEQ IDNO: 211) having a hairpin-like structure is known as a precursor of“hsa-miR-1908-5p”.

The term “hsa-miR-6857-5p gene” or “hsa-miR-6857-5p” used hereinincludes the hsa-miR-6857-5p gene (miRBase Accession No. MIMAT0027614)described in SEQ ID NO: 17, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6857-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6857” (miRBase Accession No. MI0022703, SEQ IDNO: 212) having a hairpin-like structure is known as a precursor of“hsa-miR-6857-5p”.

The term “hsa-miR-4746-3p gene” or “hsa-miR-4746-3p” used hereinincludes the hsa-miR-4746-3p gene (miRBase Accession No. MIMAT0019881)described in SEQ ID NO: 18, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-4746-3p gene can be obtainedby a method described in Persson H et al., 2011, Cancer Res, Vol. 71, p.78-86. Also, “hsa-mir-4746” (miRBase Accession No. MI0017385, SEQ ID NO:213) having a hairpin-like structure is known as a precursor of“hsa-miR-4746-3p”.

The term “hsa-miR-744-5p gene” or “hsa-miR-744-5p” used herein includesthe hsa-miR-744-5p gene (miRBase Accession No. MIMAT0004945) describedin SEQ ID NO: 19, a homolog or an ortholog of a different organismspecies, and the like. The hsa-miR-744-5p gene can be obtained by amethod described in Berezikov E et al., 2006, Genome Res, Vol. 16, p.1289-1298. Also, “hsa-mir-744” (miRBase Accession No. MI0005559, SEQ IDNO: 214) having a hairpin-like structure is known as a precursor of“hsa-miR-744-5p”.

The term “hsa-miR-4792 gene” or “hsa-miR-4792” used herein includes thehsa-miR-4792 gene (miRBase Accession No. MIMAT0019964) described in SEQID NO: 20, a homolog or an ortholog of a different organism species, andthe like. The hsa-miR-4792 gene can be obtained by a method described inPersson H et al., 2011, Cancer Res, Vol. 71, p. 78-86. Also,“hsa-mir-4792” (miRBase Accession No. MI0017439, SEQ ID NO: 215) havinga hairpin-like structure is known as a precursor of “hsa-miR-4792”.

The term “hsa-miR-564 gene” or “hsa-miR-564” used herein includes thehsa-miR-564 gene (miRBase Accession No. MIMAT0003228) described in SEQID NO: 21, a homolog or an ortholog of a different organism species, andthe like. The hsa-miR-564 gene can be obtained by a method described inCummins J M et al., 2006, Proc Natl Acad Sci USA, Vol. 103, p.3687-3692. Also, “hsa-mir-564” (miRBase Accession No. MI0003570, SEQ IDNO: 216) having a hairpin-like structure is known as a precursor of“hsa-miR-564”.

The term “hsa-miR-6791-5p gene” or “hsa-miR-6791-5p” used hereinincludes the hsa-miR-6791-5p gene (miRBase Accession No. MIMAT0027482)described in SEQ ID NO: 22, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6791-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6791” (miRBase Accession No. MI0022636, SEQ IDNO: 217) having a hairpin-like structure is known as a precursor of“hsa-miR-6791-5p”.

The term “hsa-miR-6825-5p gene” or “hsa-miR-6825-5p” used hereinincludes the hsa-miR-6825-5p gene (miRBase Accession No. MIMAT0027550)described in SEQ ID NO: 23, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6825-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6825” (miRBase Accession No. MI0022670, SEQ IDNO: 218) having a hairpin-like structure is known as a precursor of“hsa-miR-6825-5p”.

The term “hsa-miR-6826-5p gene” or “hsa-miR-6826-5p” used hereinincludes the hsa-miR-6826-5p gene (miRBase Accession No. MIMAT0027552)described in SEQ ID NO: 24, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6826-5p gene 1645. Also,“hsa-mir-6826” (miRBase Accession No. MI0022671, SEQ ID NO: 219) havinga hairpin-like structure is known as a precursor of “hsa-miR-6826-5p”.

The term “hsa-miR-4665-3p gene” or “hsa-miR-4665-3p” used hereinincludes the hsa-miR-4665-3p gene (miRBase Accession No. MIMAT0019740)described in SEQ ID NO: 25, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-4665-3p gene can be obtainedby a method described in Persson H et al., 2011, Cancer Res, Vol. 71, p.78-86. Also, “hsa-mir-4665” (miRBase Accession No. MI0017295, SEQ ID NO:220) having a hairpin-like structure is known as a precursor of“hsa-miR-4665-3p”.

The term “hsa-miR-4467 gene” or “hsa-miR-4467” used herein includes thehsa-miR-4467 gene (miRBase Accession No. MIMAT0018994) described in SEQID NO: 26, a homolog or an ortholog of a different organism species, andthe like. The hsa-miR-4467 gene can be obtained by a method described inJima D D et al., 2010, Blood, Vol. 116, e118-e127. Also, “hsa-mir-4467”(miRBase Accession No. MI0016818, SEQ ID NO: 221) having a hairpin-likestructure is known as a precursor of “hsa-miR-4467”.

The term “hsa-miR-3188 gene” or “hsa-miR-3188” used herein includes thehsa-miR-3188 gene (miRBase Accession No. MIMAT0015070) described in SEQID NO: 27, a homolog or an ortholog of a different organism species, andthe like. The hsa-miR-3188 gene can be obtained by a method described inStark M S et al., 2010, PLoS One, Vol. 5, e9685. Also, “hsa-mir-3188”(miRBase Accession No. MI0014232, SEQ ID NO: 222) having a hairpin-likestructure is known as a precursor of “hsa-miR-3188”.

The term “hsa-miR-6125 gene” or “hsa-miR-6125” used herein includes thehsa-miR-6125 gene (miRBase Accession No. MIMAT0024598) described in SEQID NO: 28, a homolog or an ortholog of a different organism species, andthe like. The hsa-miR-6125 gene can be obtained by a method described inSmith J L et al., 2012, J Virol, Vol. 86, p. 5278-5287. Also,“hsa-mir-6125” (miRBase Accession No. MI0021259, SEQ ID NO: 223) havinga hairpin-like structure is known as a precursor of “hsa-miR-6125”.

The term “hsa-miR-6756-5p gene” or “hsa-miR-6756-5p” used hereinincludes the hsa-miR-6756-5p gene (miRBase Accession No. MIMAT0027412)described in SEQ ID NO: 29, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6756-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6756” (miRBase Accession No. MI0022601, SEQ IDNO: 224) having a hairpin-like structure is known as a precursor of“hsa-miR-6756-5p”.

The term “hsa-miR-1228-3p gene” or “hsa-miR-1228-3p” used hereinincludes the hsa-miR-1228-3p gene (miRBase Accession No. MIMAT0005583)described in SEQ ID NO: 30, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-1228-3p gene can be obtainedby a method described in Berezikov E et al., 2007, Mol Cell, Vol. 28, p.328-336. Also, “hsa-mir-1228” (miRBase Accession No. MI0006318, SEQ IDNO: 225) having a hairpin-like structure is known as a precursor of“hsa-miR-1228-3p”.

The term “hsa-miR-8063 gene” or “hsa-miR-8063” used herein includes thehsa-miR-8063 gene (miRBase Accession No. MIMAT0030990) described in SEQID NO: 31, a homolog or an ortholog of a different organism species, andthe like. The hsa-miR-8063 gene can be obtained by a method described inWang H J et al., 2013, Shock, Vol. 39, p. 480-487. Also, “hsa-mir-8063”(miRBase Accession No. MI0025899, SEQ ID NO: 226) having a hairpin-likestructure is known as a precursor of “hsa-miR-8063”.

The term “hsa-miR-8069 gene” or “hsa-miR-8069” used herein includes thehsa-miR-8069 gene (miRBase Accession No. MIMAT0030996) described in SEQID NO: 32, a homolog or an ortholog of a different organism species, andthe like. The hsa-miR-8069 gene can be obtained by a method described inWang H J et al., 2013, Shock, Vol. 39, p. 480-487. Also, “hsa-mir-8069”(miRBase Accession No. MI0025905, SEQ ID NO: 227) having a hairpin-likestructure is known as a precursor of “hsa-miR-8069”.

The term “hsa-miR-6875-5p gene” or “hsa-miR-6875-5p” used hereinincludes the hsa-miR-6875-5p gene (miRBase Accession No. MIMAT0027650)described in SEQ ID NO: 33, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6875-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6875” (miRBase Accession No. MI0022722, SEQ IDNO: 228) having a hairpin-like structure is known as a precursor of“hsa-miR-6875-5p”.

The term “hsa-miR-3185 gene” or “hsa-miR-3185” used herein includes thehsa-miR-3185 gene (miRBase Accession No. MIMAT0015065) described in SEQID NO: 34, a homolog or an ortholog of a different organism species, andthe like. The hsa-miR-3185 gene can be obtained by a method described inStark M S et al., 2010, PLoS One, Vol. 5, e9685. Also, “hsa-mir-3185”(miRBase Accession No. MI0014227, SEQ ID NO: 229) having a hairpin-likestructure is known as a precursor of “hsa-miR-3185”.

The term “hsa-miR-4433b-3p gene” or “hsa-miR-4433b-3p” used hereinincludes the hsa-miR-4433b-3p gene (miRBase Accession No. MIMAT0030414)described in SEQ ID NO: 35, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-4433b-3p gene can beobtained by a method described in Ple H et al., 2012, PLoS One, Vol. 7,e50746. Also, “hsa-mir-4433b” (miRBase Accession No. MI0025511, SEQ IDNO: 230) having a hairpin-like structure is known as a precursor of“hsa-miR-4433b-3p”.

The term “hsa-miR-6887-5p gene” or “hsa-miR-6887-5p” used hereinincludes the hsa-miR-6887-5p gene (miRBase Accession No. MIMAT0027674)described in SEQ ID NO: 36, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6887-5p gene 1645. Also,“hsa-mir-6887” (miRBase Accession No. MI0022734, SEQ ID NO: 231) havinga hairpin-like structure is known as a precursor of “hsa-miR-6887-5p”.

The term “hsa-miR-128-1-5p gene” or “hsa-miR-128-1-5p” used hereinincludes the hsa-miR-128-1-5p gene (miRBase Accession No. MIMAT0026477)described in SEQ ID NO: 37, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-128-1-5p gene can beobtained by a method described in Lagos-Quintana M et al., 2002, CurrBiol, Vol. 12, p. 735-739. Also, “hsa-mir-128-1” (miRBase Accession No.MI0000447, SEQ ID NO: 232) having a hairpin-like structure is known as aprecursor of “hsa-miR-128-1-5p”.

The term “hsa-miR-6724-5p gene” or “hsa-miR-6724-5p” used hereinincludes the hsa-miR-6724-5p gene (miRBase Accession No. MIMAT0025856)described in SEQ ID NO: 38, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6724-5p gene can be obtainedby a method described in Li Y et al., 2012, Gene, Vol. 497, p. 330-335.Also, “hsa-mir-6724” (miRBase Accession No. MI0022559, SEQ ID NO: 233)having a hairpin-like structure is known as a precursor of“hsa-miR-6724-5p”.

The term “hsa-miR-1914-3p gene” or “hsa-miR-1914-3p” used hereinincludes the hsa-miR-1914-3p gene (miRBase Accession No. MIMAT0007890)described in SEQ ID NO: 39, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-1914-3p gene can be obtainedby a method described in Bar M et al., 2008, Stem Cells, Vol. 26, p.2496-2505. Also, “hsa-mir-1914” (miRBase Accession No. MI0008335, SEQ IDNO: 234) having a hairpin-like structure is known as a precursor of“hsa-miR-1914-3p”.

The term “hsa-miR-1225-5p gene” or “hsa-miR-1225-5p” used hereinincludes the hsa-miR-1225-5p gene (miRBase Accession No. MIMAT0005572)described in SEQ ID NO: 40, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-1225-5p gene can be obtainedby a method described in Berezikov E et al., 2007, Mol Cell, Vol. 28, p.328-336. Also, “hsa-mir-1225” (miRBase Accession No. MI0006311, SEQ IDNO: 235) having a hairpin-like structure is known as a precursor of“hsa-miR-1225-5p”.

The term “hsa-miR-4419b gene” or “hsa-miR-4419b” used herein includesthe hsa-miR-4419b gene (miRBase Accession No. MIMAT0019034) described inSEQ ID NO: 41, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-4419b gene can be obtained by a methoddescribed in Jima D D et al., 2010, Blood, Vol. 116, e118-e127. Also,“hsa-mir-4419b” (miRBase Accession No. MI0016861, SEQ ID NO: 236) havinga hairpin-like structure is known as a precursor of “hsa-miR-4419b”.

The term “hsa-miR-7110-5p gene” or “hsa-miR-7110-5p” used hereinincludes the hsa-miR-7110-5p gene (miRBase Accession No. MIMAT0028117)described in SEQ ID NO: 42, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-7110-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-7110” (miRBase Accession No. MI0022961, SEQ IDNO: 237) having a hairpin-like structure is known as a precursor of“hsa-miR-7110-5p”.

The term “hsa-miR-187-5p gene” or “hsa-miR-187-5p” used herein includesthe hsa-miR-187-5p gene (miRBase Accession No. MIMAT0004561) describedin SEQ ID NO: 43, a homolog or an ortholog of a different organismspecies, and the like. The hsa-miR-187-5p gene can be obtained by amethod described in Lim L P et al., 2003, Science, Vol. 299, p. 1540.Also, “hsa-mir-187” (miRBase Accession No. MI0000274, SEQ ID NO: 238)having a hairpin-like structure is known as a precursor of“hsa-miR-187-5p”.

The term “hsa-miR-3184-5p gene” or “hsa-miR-3184-5p” used hereinincludes the hsa-miR-3184-5p gene (miRBase Accession No. MIMAT0015064)described in SEQ ID NO: 44, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-3184-5p gene can be obtainedby a method described in Stark M S et al., 2010, PLoS One, Vol. 5,e9685. Also, “hsa-mir-3184” (miRBase Accession No. MI0014226, SEQ ID NO:239) having a hairpin-like structure is known as a precursor of“hsa-miR-3184-5p”.

The term “hsa-miR-204-3p gene” or “hsa-miR-204-3p” used herein includesthe hsa-miR-204-3p gene (miRBase Accession No. MIMAT0022693) describedin SEQ ID NO: 45, a homolog or an ortholog of a different organismspecies, and the like. The hsa-miR-204-3p gene can be obtained by amethod described in Lim L P et al., 2003, Science, Vol. 299, p. 1540.Also, “hsa-mir-204” (miRBase Accession No. MI0000284, SEQ ID NO: 240)having a hairpin-like structure is known as a precursor of“hsa-miR-204-3p”.

The term “hsa-miR-5572 gene” or “hsa-miR-5572” used herein includes thehsa-miR-5572 gene (miRBase Accession No. MIMAT0022260) described in SEQID NO: 46, a homolog or an ortholog of a different organism species, andthe like. The hsa-miR-5572 gene can be obtained by a method described inTandon Metal., 2012, Oral Dis, Vol. 18, p. 127-131. Also, “hsa-mir-5572”(miRBase Accession No. MI0019117, SEQ ID NO: 241) having a hairpin-likestructure is known as a precursor of “hsa-miR-5572”.

The term “hsa-miR-6729-5p gene” or “hsa-miR-6729-5p” used hereinincludes the hsa-miR-6729-5p gene (miRBase Accession No. MIMAT0027359)described in SEQ ID NO: 47, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6729-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6729” (miRBase Accession No. MI0022574, SEQ IDNO: 242) having a hairpin-like structure is known as a precursor of“hsa-miR-6729-5p”.

The term “hsa-miR-615-5p gene” or “hsa-miR-615-5p” used herein includesthe hsa-miR-615-5p gene (miRBase Accession No. MIMAT0004804) describedin SEQ ID NO: 48, a homolog or an ortholog of a different organismspecies, and the like. The hsa-miR-615-5p gene can be obtained by amethod described in Cummins J M et al., 2006, Proc Natl Acad Sci USA,Vol. 103, p. 3687-3692. Also, “hsa-mir-615” (miRBase Accession No.MI0003628, SEQ ID NO: 243) having a hairpin-like structure is known as aprecursor of “hsa-miR-615-5p”.

The term “hsa-miR-6749-5p gene” or “hsa-miR-6749-5p” used hereinincludes the hsa-miR-6749-5p gene (miRBase Accession No. MIMAT0027398)described in SEQ ID NO: 49, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6749-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6749” (miRBase Accession No. MI0022594, SEQ IDNO: 244) having a hairpin-like structure is known as a precursor of“hsa-miR-6749-5p”.

The term “hsa-miR-6515-3p gene” or “hsa-miR-6515-3p” used hereinincludes the hsa-miR-6515-3p gene (miRBase Accession No. MIMAT0025487)described in SEQ ID NO: 50, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6515-3p gene can be obtainedby a method described in Joyce C E et al., 2011, Hum Mol Genet, Vol. 20,p. 4025-4040. Also, “hsa-mir-6515” (miRBase Accession No. MI0022227, SEQID NO: 245) having a hairpin-like structure is known as a precursor of“hsa-miR-6515-3p”.

The term “hsa-miR-3937 gene” or “hsa-miR-3937” used herein includes thehsa-miR-3937 gene (miRBase Accession No. MIMAT0018352) described in SEQID NO: 51, a homolog or an ortholog of a different organism species, andthe like. The hsa-miR-3937 gene can be obtained by a method described inLiao J Y et al., 2010, PLoS One, Vol. 5, e10563. Also, “hsa-mir-3937”(miRBase Accession No. MI0016593, SEQ ID NO: 246) having a hairpin-likestructure is known as a precursor of “hsa-miR-3937”.

The term “hsa-miR-6840-3p gene” or “hsa-miR-6840-3p” used hereinincludes the hsa-miR-6840-3p gene (miRBase Accession No. MIMAT0027583)described in SEQ ID NO: 52, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6840-3p gene 1645. Also,“hsa-mir-6840” (miRBase Accession No. MI0022686, SEQ ID NO: 247) havinga hairpin-like structure is known as a precursor of “hsa-miR-6840-3p”.

The term “hsa-miR-6893-5p gene” or “hsa-miR-6893-5p” used hereinincludes the hsa-miR-6893-5p gene (miRBase Accession No. MIMAT0027686)described in SEQ ID NO: 53, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6893-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6893” (miRBase Accession No. MI0022740, SEQ IDNO: 248) having a hairpin-like structure is known as a precursor of“hsa-miR-6893-5p”.

The term “hsa-miR-4728-5p gene” or “hsa-miR-4728-5p” used hereinincludes the hsa-miR-4728-5p gene (miRBase Accession No. MIMAT0019849)described in SEQ ID NO: 54, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-4728-5p gene can be obtainedby a method described in Persson H et al., 2011, Cancer Res, Vol. 71, p.78-86. Also, “hsa-mir-4728” (miRBase Accession No. MI0017365, SEQ ID NO:249) having a hairpin-like structure is known as a precursor of“hsa-miR-4728-5p”.

The term “hsa-miR-6717-5p gene” or “hsa-miR-6717-5p” used hereinincludes the hsa-miR-6717-5p gene (miRBase Accession No. MIMAT0025846)described in SEQ ID NO: 55, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6717-5p gene can be obtainedby a method described in Li Y et al., 2012, Gene, Vol. 497, p. 330-335.Also, “hsa-mir-6717” (miRBase Accession No. MI0022551, SEQ ID NO: 250)having a hairpin-like structure is known as a precursor of“hsa-miR-6717-5p”.

The term “hsa-miR-7113-3p gene” or “hsa-miR-7113-3p” used hereinincludes the hsa-miR-7113-3p gene (miRBase Accession No. MIMAT0028124)described in SEQ ID NO: 56, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-7113-3p gene 1645. Also,“hsa-mir-7113” (miRBase Accession No. MI0022964, SEQ ID NO: 251) havinga hairpin-like structure is known as a precursor of “hsa-miR-7113-3p”.

The term “hsa-miR-4665-5p gene” or “hsa-miR-4665-5p” used hereinincludes the hsa-miR-4665-5p gene (miRBase Accession No. MIMAT0019739)described in SEQ ID NO: 57, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-4665-5p gene can be obtainedby a method described in Persson H et al., 2011, Cancer Res, Vol. 71, p.78-86. Also, “hsa-mir-4665” (miRBase Accession No. MI0017295, SEQ ID NO:220) having a hairpin-like structure is known as a precursor of“hsa-miR-4665-5p”.

The term “hsa-miR-642b-3p gene” or “hsa-miR-642b-3p” used hereinincludes the hsa-miR-642b-3p gene (miRBase Accession No. MIMAT0018444)described in SEQ ID NO: 58, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-642b-3p gene can be obtainedby a method described in Witten D et al., 2010, BMC Biol, Vol. 8, p. 58.Also, “hsa-mir-642b” (miRBase Accession No. MI0016685, SEQ ID NO: 252)having a hairpin-like structure is known as a precursor of“hsa-miR-642b-3p”.

The term “hsa-miR-7109-5p gene” or “hsa-miR-7109-5p” used hereinincludes the hsa-miR-7109-5p gene (miRBase Accession No. MIMAT0028115)described in SEQ ID NO: 59, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-7109-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-7109” (miRBase Accession No. MI0022960, SEQ IDNO: 253) having a hairpin-like structure is known as a precursor of“hsa-miR-7109-5p”.

The term “hsa-miR-6842-5p gene” or “hsa-miR-6842-5p” used hereinincludes the hsa-miR-6842-5p gene (miRBase Accession No. MIMAT0027586)described in SEQ ID NO: 60, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6842-5p gene 1645. Also,“hsa-mir-6842” (miRBase Accession No. MI0022688, SEQ ID NO: 254) havinga hairpin-like structure is known as a precursor of “hsa-miR-6842-5p”.

The term “hsa-miR-4442 gene” or “hsa-miR-4442” used herein includes thehsa-miR-4442 gene (miRBase Accession No. MIMAT0018960) described in SEQID NO: 61, a homolog or an ortholog of a different organism species, andthe like. The hsa-miR-4442 gene can be obtained by a method described inJima D D et al., 2010, Blood, Vol. 116, e118-e127. Also, “hsa-mir-4442”(miRBase Accession No. MI0016785, SEQ ID NO: 255) having a hairpin-likestructure is known as a precursor of “hsa-miR-4442”.

The term “hsa-miR-4433-3p gene” or “hsa-miR-4433-3p” used hereinincludes the hsa-miR-4433-3p gene (miRBase Accession No. MIMAT0018949)described in SEQ ID NO: 62, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-4433-3p gene can be obtainedby a method described in Jima D D et al., 2010, Blood, Vol. 116,e118-e127. Also, “hsa-mir-4433” (miRBase Accession No. MI0016773, SEQ IDNO: 256) having a hairpin-like structure is known as a precursor of“hsa-miR-4433-3p”.

The term “hsa-miR-4707-5p gene” or “hsa-miR-4707-5p” used hereinincludes the hsa-miR-4707-5p gene (miRBase Accession No. MIMAT0019807)described in SEQ ID NO: 63, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-4707-5p gene can be obtainedby a method described in Persson H et al., 2011, Cancer Res, Vol. 71, p.78-86. Also, “hsa-mir-4707” (miRBase Accession No. MI0017340, SEQ ID NO:257) having a hairpin-like structure is known as a precursor of“hsa-miR-4707-5p”.

The term “hsa-miR-6126 gene” or “hsa-miR-6126” used herein includes thehsa-miR-6126 gene (miRBase Accession No. MIMAT0024599) described in SEQID NO: 64, a homolog or an ortholog of a different organism species, andthe like. The hsa-miR-6126 gene can be obtained by a method described inSmith J L et al., 2012, J Virol, Vol. 86, p. 5278-5287. Also,“hsa-mir-6126” (miRBase Accession No. MI0021260, SEQ ID NO: 258) havinga hairpin-like structure is known as a precursor of “hsa-miR-6126”.

The term “hsa-miR-4449 gene” or “hsa-miR-4449” used herein includes thehsa-miR-4449 gene (miRBase Accession No. MIMAT0018968) described in SEQID NO: 65, a homolog or an ortholog of a different organism species, andthe like. The hsa-miR-4449 gene can be obtained by a method described inJima D D et al., 2010, Blood, Vol. 116, e118-e127. Also, “hsa-mir-4449”(miRBase Accession No. MI0016792, SEQ ID NO: 259) having a hairpin-likestructure is known as a precursor of “hsa-miR-4449”.

The term “hsa-miR-4706 gene” or “hsa-miR-4706” used herein includes thehsa-miR-4706 gene (miRBase Accession No. MIMAT0019806) described in SEQID NO: 66, a homolog or an ortholog of a different organism species, andthe like. The hsa-miR-4706 gene can be obtained by a method described inPersson H et al., 2011, Cancer Res, Vol. 71, p. 78-86. Also,“hsa-mir-4706” (miRBase Accession No. MI0017339, SEQ ID NO: 260) havinga hairpin-like structure is known as a precursor of “hsa-miR-4706”.

The term “hsa-miR-1913 gene” or “hsa-miR-1913” used herein includes thehsa-miR-1913 gene (miRBase Accession No. MIMAT0007888) described in SEQID NO: 67, a homolog or an ortholog of a different organism species, andthe like. The hsa-miR-1913 gene can be obtained by a method described inBar M et al., 2008, Stem Cells, Vol. 26, p. 2496-2505. Also,“hsa-mir-1913” (miRBase Accession No. MI0008334, SEQ ID NO: 261) havinga hairpin-like structure is known as a precursor of “hsa-miR-1913”.

The term “hsa-miR-602 gene” or “hsa-miR-602” used herein includes thehsa-miR-602 gene (miRBase Accession No. MIMAT0003270) described in SEQID NO: 68, a homolog or an ortholog of a different organism species, andthe like. The hsa-miR-602 gene can be obtained by a method described inCummins J M et al., 2006, Proc Natl Acad Sci USA, Vol. 103, p.3687-3692. Also, “hsa-mir-602” (miRBase Accession No. MI0003615, SEQ IDNO: 262) having a hairpin-like structure is known as a precursor of“hsa-miR-602”.

The term “hsa-miR-939-5p gene” or “hsa-miR-939-5p” used herein includesthe hsa-miR-939-5p gene (miRBase Accession No. MIMAT0004982) describedin SEQ ID NO: 69, a homolog or an ortholog of a different organismspecies, and the like. The hsa-miR-939-5p gene can be obtained by amethod described in Lui W O et al., 2007, Cancer Res, Vol. 67, p.6031-6043. Also, “hsa-mir-939” (miRBase Accession No. MI0005761, SEQ IDNO: 263) having a hairpin-like structure is known as a precursor of“hsa-miR-939-5p”.

The term “hsa-miR-4695-5p gene” or “hsa-miR-4695-5p” used hereinincludes the hsa-miR-4695-5p gene (miRBase Accession No. MIMAT0019788)described in SEQ ID NO: 70, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-4695-5p gene can be obtainedby a method described in Persson H et al., 2011, Cancer Res, Vol. 71, p.78-86. Also, “hsa-mir-4695” (miRBase Accession No. MI0017328, SEQ ID NO:264) having a hairpin-like structure is known as a precursor of“hsa-miR-4695-5p”.

The term “hsa-miR-711 gene” or “hsa-miR-711” used herein includes thehsa-miR-711 gene (miRBase Accession No. MIMAT0012734) described in SEQID NO: 71, a homolog or an ortholog of a different organism species, andthe like. The hsa-miR-711 gene can be obtained by a method described inArtzi S et al., 2008, BMC Bioinformatics, Vol. 9, p. 39. Also,“hsa-mir-711” (miRBase Accession No. MI0012488, SEQ ID NO: 265) having ahairpin-like structure is known as a precursor of “hsa-miR-711”.

The term “hsa-miR-6816-5p gene” or “hsa-miR-6816-5p” used hereinincludes the hsa-miR-6816-5p gene (miRBase Accession No. MIMAT0027532)described in SEQ ID NO: 72, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6816-5p gene 1645. Also,“hsa-mir-6816” (miRBase Accession No. MI0022661, SEQ ID NO: 266) havinga hairpin-like structure is known as a precursor of “hsa-miR-6816-5p”.

The term “hsa-miR-4632-5p gene” or “hsa-miR-4632-5p” used hereinincludes the hsa-miR-4632-5p gene (miRBase Accession No. MIMAT0022977)described in SEQ ID NO: 73, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-4632-5p gene can be obtainedby a method described in Persson H et al., 2011, Cancer Res, Vol. 71, p.78-86. Also, “hsa-mir-4632” (miRBase Accession No. MI0017259, SEQ ID NO:267) having a hairpin-like structure is known as a precursor of“hsa-miR-4632-5p”.

The term “hsa-miR-6721-5p gene” or “hsa-miR-6721-5p” used hereinincludes the hsa-miR-6721-5p gene (miRBase Accession No. MIMAT0025852)described in SEQ ID NO: 74, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6721-5p gene can be obtainedby a method described in Li Y et al., 2012, Gene, Vol. 497, p. 330-335.Also, “hsa-mir-6721” (miRBase Accession No. MI0022556, SEQ ID NO: 268)having a hairpin-like structure is known as a precursor of“hsa-miR-6721-5p”.

The term “hsa-miR-7847-3p gene” or “hsa-miR-7847-3p” used hereinincludes the hsa-miR-7847-3p gene (miRBase Accession No. MIMAT0030422)described in SEQ ID NO: 75, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-7847-3p gene can be obtainedby a method described in Ple H et al., 2012, PLoS One, Vol. 7, e50746.Also, “hsa-mir-7847” (miRBase Accession No. MI0025517, SEQ ID NO: 269)having a hairpin-like structure is known as a precursor of“hsa-miR-7847-3p”.

The term “hsa-miR-6132 gene” or “hsa-miR-6132” used herein includes thehsa-miR-6132 gene (miRBase Accession No. MIMAT0024616) described in SEQID NO: 76, a homolog or an ortholog of a different organism species, andthe like. The hsa-miR-6132 gene can be obtained by a method described inDannemann M et al., 2012, Genome Biol Evol, Vol. 4, p. 552-564. Also,“hsa-mir-6132” (miRBase Accession No. MI0021277, SEQ ID NO: 270) havinga hairpin-like structure is known as a precursor of “hsa-miR-6132”.

The term “hsa-miR-887-3p gene” or “hsa-miR-887-3p” used herein includesthe hsa-miR-887-3p gene (miRBase Accession No. MIMAT0004951) describedin SEQ ID NO: 77, a homolog or an ortholog of a different organismspecies, and the like. The hsa-miR-887-3p gene can be obtained by amethod described in Berezikov E et al., 2006, Genome Res, Vol. 16, p.1289-1298. Also, “hsa-mir-887” (miRBase Accession No. MI0005562, SEQ IDNO: 271) having a hairpin-like structure is known as a precursor of“hsa-miR-887-3p”.

The term “hsa-miR-3679-3p gene” or “hsa-miR-3679-3p” used hereinincludes the hsa-miR-3679-3p gene (miRBase Accession No. MIMAT0018105)described in SEQ ID NO: 78, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-3679-3p gene can be obtainedby a method described in Creighton C J et al., 2010, PLoS One, Vol. 5,e9637. Also, “hsa-mir-3679” (miRBase Accession No. MI0016080, SEQ ID NO:205) having a hairpin-like structure is known as a precursor of“hsa-miR-3679-3p”.

The term “hsa-miR-6784-5p gene” or “hsa-miR-6784-5p” used hereinincludes the hsa-miR-6784-5p gene (miRBase Accession No. MIMAT0027468)described in SEQ ID NO: 79, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6784-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6784” (miRBase Accession No. MI0022629, SEQ IDNO: 272) having a hairpin-like structure is known as a precursor of“hsa-miR-6784-5p”.

The term “hsa-miR-1249 gene” or “hsa-miR-1249” used herein includes thehsa-miR-1249 gene (miRBase Accession No. MIMAT0005901) described in SEQID NO: 80, a homolog or an ortholog of a different organism species, andthe like. The hsa-miR-1249 gene can be obtained by a method described inMorin R D et al., 2008, Genome Res, Vol. 18, p. 610-621. Also,“hsa-mir-1249” (miRBase Accession No. MI0006384, SEQ ID NO: 273) havinga hairpin-like structure is known as a precursor of “hsa-miR-1249”.

The term “hsa-miR-937-5p gene” or “hsa-miR-937-5p” used herein includesthe hsa-miR-937-5p gene (miRBase Accession No. MIMAT0022938) describedin SEQ ID NO: 81, a homolog or an ortholog of a different organismspecies, and the like. The hsa-miR-937-5p gene can be obtained by amethod described in Lui W O et al., 2007, Cancer Res, Vol. 67, p.6031-6043. Also, “hsa-mir-937” (miRBase Accession No. MI0005759, SEQ IDNO: 274) having a hairpin-like structure is known as a precursor of“hsa-miR-937-5p”.

The term “hsa-miR-5195-3p gene” or “hsa-miR-5195-3p” used hereinincludes the hsa-miR-5195-3p gene (miRBase Accession No. MIMAT0021127)described in SEQ ID NO: 82, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-5195-3p gene can be obtainedby a method described in Schotte D et al., 2011, Leukemia, Vol. 25, p.1389-1399. Also, “hsa-mir-5195” (miRBase Accession No. MI0018174, SEQ IDNO: 275) having a hairpin-like structure is known as a precursor of“hsa-miR-5195-3p”.

The term “hsa-miR-6732-5p gene” or “hsa-miR-6732-5p” used hereinincludes the hsa-miR-6732-5p gene (miRBase Accession No. MIMAT0027365)described in SEQ ID NO: 83, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6732-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6732” (miRBase Accession No. MI0022577, SEQ IDNO: 276) having a hairpin-like structure is known as a precursor of“hsa-miR-6732-5p”.

The term “hsa-miR-4417 gene” or “hsa-miR-4417” used herein includes thehsa-miR-4417 gene (miRBase Accession No. MIMAT0018929) described in SEQID NO: 84, a homolog or an ortholog of a different organism species, andthe like. The hsa-miR-4417 gene can be obtained by a method described inJima D D et al., 2010, Blood, Vol. 116, e118-e127. Also, “hsa-mir-4417”(miRBase Accession No. MI0016753, SEQ ID NO: 277) having a hairpin-likestructure is known as a precursor of “hsa-miR-4417”.

The term “hsa-miR-4281 gene” or “hsa-miR-4281” used herein includes thehsa-miR-4281 gene (miRBase Accession No. MIMAT0016907) described in SEQID NO: 85, a homolog or an ortholog of a different organism species, andthe like. The hsa-miR-4281 gene can be obtained by a method described inGoff L A et al., 2009, PLoS One, Vol. 4, e7192. Also, “hsa-mir-4281”(miRBase Accession No. MI0015885, SEQ ID NO: 278) having a hairpin-likestructure is known as a precursor of “hsa-miR-4281”.

The term “hsa-miR-4734 gene” or “hsa-miR-4734” used herein includes thehsa-miR-4734 gene (miRBase Accession No. MIMAT0019859) described in SEQID NO: 86, a homolog or an ortholog of a different organism species, andthe like. The hsa-miR-4734 gene can be obtained by a method described inPersson H et al., 2011, Cancer Res, Vol. 71, p. 78-86. Also,“hsa-mir-4734” (miRBase Accession No. MI0017371, SEQ ID NO: 279) havinga hairpin-like structure is known as a precursor of “hsa-miR-4734”.

The term “hsa-miR-6766-3p gene” or “hsa-miR-6766-3p” used hereinincludes the hsa-miR-6766-3p gene (miRBase Accession No. MIMAT0027433)described in SEQ ID NO: 87, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6766-3p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6766” (miRBase Accession No. MI0022611, SEQ IDNO: 280) having a hairpin-like structure is known as a precursor of“hsa-miR-6766-3p”.

The term “hsa-miR-663a gene” or “hsa-miR-663a” used herein includes thehsa-miR-663a gene (miRBase Accession No. MIMAT0003326) described in SEQID NO: 88, a homolog or an ortholog of a different organism species, andthe like. The hsa-miR-663a gene can be obtained by a method described inCummins J M et al., 2006, Proc Natl Acad Sci USA, Vol. 103, p.3687-3692. Also, “hsa-mir-663a” (miRBase Accession No. MI0003672, SEQ IDNO: 281) having a hairpin-like structure is known as a precursor of“hsa-miR-663a”.

The term “hsa-miR-4513 gene” or “hsa-miR-4513” used herein includes thehsa-miR-4513 gene (miRBase Accession No. MIMAT0019050) described in SEQID NO: 89, a homolog or an ortholog of a different organism species, andthe like. The hsa-miR-4513 gene can be obtained by a method described inJima D D et al., 2010, Blood, Vol. 116, e118-e127. Also, “hsa-mir-4513”(miRBase Accession No. MI0016879, SEQ ID NO: 282) having a hairpin-likestructure is known as a precursor of “hsa-miR-4513”.

The term “hsa-miR-6781-5p gene” or “hsa-miR-6781-5p” used hereinincludes the hsa-miR-6781-5p gene (miRBase Accession No. MIMAT0027462)described in SEQ ID NO: 90, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6781-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6781” (miRBase Accession No. MI0022626, SEQ IDNO: 283) having a hairpin-like structure is known as a precursor of“hsa-miR-6781-5p”.

The term “hsa-miR-1227-5p gene” or “hsa-miR-1227-5p” used hereinincludes the hsa-miR-1227-5p gene (miRBase Accession No. MIMAT0022941)described in SEQ ID NO: 91, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-1227-5p gene can be obtainedby a method described in Berezikov E et al., 2007, Mol Cell, Vol. 28, p.328-336. Also, “hsa-mir-1227” (miRBase Accession No. MI0006316, SEQ IDNO: 284) having a hairpin-like structure is known as a precursor of“hsa-miR-1227-5p”.

The term “hsa-miR-6845-5p gene” or “hsa-miR-6845-5p” used hereinincludes the hsa-miR-6845-5p gene (miRBase Accession No. MIMAT0027590)described in SEQ ID NO: 92, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6845-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6845” (miRBase Accession No. MI0022691, SEQ IDNO: 285) having a hairpin-like structure is known as a precursor of“hsa-miR-6845-5p”.

The term “hsa-miR-6798-5p gene” or “hsa-miR-6798-5p” used hereinincludes the hsa-miR-6798-5p gene (miRBase Accession No. MIMAT0027496)described in SEQ ID NO: 93, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6798-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6798” (miRBase Accession No. MI0022643, SEQ IDNO: 286) having a hairpin-like structure is known as a precursor of“hsa-miR-6798-5p”.

The term “hsa-miR-3620-5p gene” or “hsa-miR-3620-5p” used hereinincludes the hsa-miR-3620-5p gene (miRBase Accession No. MIMAT0022967)described in SEQ ID NO: 94, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-3620-5p gene can be obtainedby a method described in Witten D et al., 2010, BMC Biol, Vol. 8, p. 58.Also, “hsa-mir-3620” (miRBase Accession No. MI0016011, SEQ ID NO: 287)having a hairpin-like structure is known as a precursor of“hsa-miR-3620-5p”.

The term “hsa-miR-1915-5p gene” or “hsa-miR-1915-5p” used hereinincludes the hsa-miR-1915-5p gene (miRBase Accession No. MIMAT0007891)described in SEQ ID NO: 95, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-1915-5p gene can be obtainedby a method described in Bar M et al., 2008, Stem Cells, Vol. 26, p.2496-2505. Also, “hsa-mir-1915” (miRBase Accession No. MI0008336, SEQ IDNO: 288) having a hairpin-like structure is known as a precursor of“hsa-miR-1915-5p”.

The term “hsa-miR-4294 gene” or “hsa-miR-4294” used herein includes thehsa-miR-4294 gene (miRBase Accession No. MIMAT0016849) described in SEQID NO: 96, a homolog or an ortholog of a different organism species, andthe like. The hsa-miR-4294 gene can be obtained by a method described inGoff L A et al., 2009, PLoS One, Vol. 4, e7192. Also, “hsa-mir-4294”(miRBase Accession No. MI0015827, SEQ ID NO: 289) having a hairpin-likestructure is known as a precursor of “hsa-miR-4294”.

The term “hsa-miR-642a-3p gene” or “hsa-miR-642a-3p” used hereinincludes the hsa-miR-642a-3p gene (miRBase Accession No. MIMAT0020924)described in SEQ ID NO: 97, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-642a-3p gene can be obtainedby a method described in Cummins J M et al., 2006, Proc Natl Acad SciUSA, Vol. 103, p. 3687-3692. Also, “hsa-mir-642a” (miRBase Accession No.MI0003657, SEQ ID NO: 290) having a hairpin-like structure is known as aprecursor of “hsa-miR-642a-3p”.

The term “hsa-miR-371a-5p gene” or “hsa-miR-371a-5p” used hereinincludes the hsa-miR-371a-5p gene (miRBase Accession No. MIMAT0004687)described in SEQ ID NO: 98, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-371a-5p gene can be obtainedby a method described in Suh M R et al., 2004, Dev Biol, Vol. 270, p.488-498. Also, “hsa-mir-371a” (miRBase Accession No. MI0000779, SEQ IDNO: 291) having a hairpin-like structure is known as a precursor of“hsa-miR-371a-5p”.

The term “hsa-miR-940 gene” or “hsa-miR-940” used herein includes thehsa-miR-940 gene (miRBase Accession No. MIMAT0004983) described in SEQID NO: 99, a homolog or an ortholog of a different organism species, andthe like. The hsa-miR-940 gene can be obtained by a method described inLui W O et al., 2007, Cancer Res, Vol. 67, p. 6031-6043. Also,“hsa-mir-940” (miRBase Accession No. MI0005762, SEQ ID NO: 292) having ahairpin-like structure is known as a precursor of “hsa-miR-940”.

The term “hsa-miR-4450 gene” or “hsa-miR-4450” used herein includes thehsa-miR-4450 gene (miRBase Accession No. MIMAT0018971) described in SEQID NO: 100, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-4450 gene can be obtained by a methoddescribed in Jima D D et al., 2010, Blood, Vol. 116, e118-e127. Also,“hsa-mir-4450” (miRBase Accession No. MI0016795, SEQ ID NO: 293) havinga hairpin-like structure is known as a precursor of “hsa-miR-4450”.

The term “hsa-miR-4723-5p gene” or “hsa-miR-4723-5p” used hereinincludes the hsa-miR-4723-5p gene (miRBase Accession No. MIMAT0019838)described in SEQ ID NO: 101, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-4723-5p gene can be obtainedby a method described in Persson H et al., 2011, Cancer Res, Vol. 71, p.78-86. Also, “hsa-mir-4723” (miRBase Accession No. MI0017359, SEQ ID NO:294) having a hairpin-like structure is known as a precursor of“hsa-miR-4723-5p”.

The term “hsa-miR-1469 gene” or “hsa-miR-1469” used herein includes thehsa-miR-1469 gene (miRBase Accession No. MIMAT0007347) described in SEQID NO: 102, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-1469 gene can be obtained by a methoddescribed in Kawaji H et al., 2008, BMC Genomics, Vol. 9, p. 157. Also,“hsa-mir-1469” (miRBase Accession No. MI0007074, SEQ ID NO: 295) havinga hairpin-like structure is known as a precursor of “hsa-miR-1469”.

The term “hsa-miR-6861-5p gene” or “hsa-miR-6861-5p” used hereinincludes the hsa-miR-6861-5p gene (miRBase Accession No. MIMAT0027623)described in SEQ ID NO: 103, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6861-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6861” (miRBase Accession No. MI0022708, SEQ IDNO: 296) having a hairpin-like structure is known as a precursor of“hsa-miR-6861-5p”.

The term “hsa-miR-7975 gene” or “hsa-miR-7975” used herein includes thehsa-miR-7975 gene (miRBase Accession No. MIMAT0031178) described in SEQID NO: 104, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-7975 gene can be obtained by a methoddescribed in Velthut-Meikas A et al., 2013, Mol Endocrinol, online.Also, “hsa-mir-7975” (miRBase Accession No. MI0025751, SEQ ID NO: 297)having a hairpin-like structure is known as a precursor of“hsa-miR-7975”.

The term “hsa-miR-6879-5p gene” or “hsa-miR-6879-5p” used hereinincludes the hsa-miR-6879-5p gene (miRBase Accession No. MIMAT0027658)described in SEQ ID NO: 105, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6879-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6879” (miRBase Accession No. MI0022726, SEQ IDNO: 298) having a hairpin-like structure is known as a precursor of“hsa-miR-6879-5p”.

The term “hsa-miR-6802-5p gene” or “hsa-miR-6802-5p” used hereinincludes the hsa-miR-6802-5p gene (miRBase Accession No. MIMAT0027504)described in SEQ ID NO: 106, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6802-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6802” (miRBase Accession No. MI0022647, SEQ IDNO: 299) having a hairpin-like structure is known as a precursor of“hsa-miR-6802-5p”.

The term “hsa-miR-1268b gene” or “hsa-miR-1268b” used herein includesthe hsa-miR-1268b gene (miRBase Accession No. MIMAT0018925) described inSEQ ID NO: 107, a homolog or an ortholog of a different organismspecies, and the like. The hsa-miR-1268b gene can be obtained by amethod described in Jima D D et al., 2010, Blood, Vol. 116, e118-e127.Also, “hsa-mir-1268b” (miRBase Accession No. MI0016748, SEQ ID NO: 300)having a hairpin-like structure is known as a precursor of“hsa-miR-1268b”.

The term “hsa-miR-663b gene” or “hsa-miR-663b” used herein includes thehsa-miR-663b gene (miRBase Accession No. MIMAT0005867) described in SEQID NO: 108, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-663b gene can be obtained by a methoddescribed in Takada S et al., 2008, Leukemia, Vol. 22, p. 1274-1278.Also, “hsa-mir-663b” (miRBase Accession No. MI0006336, SEQ ID NO: 301)having a hairpin-like structure is known as a precursor of“hsa-miR-663b”.

The term “hsa-miR-125a-3p gene” or “hsa-miR-125a-3p” used hereinincludes the hsa-miR-125a-3p gene (miRBase Accession No. MIMAT0004602)described in SEQ ID NO: 109, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-125a-3p gene can be obtainedby a method described in Lagos-Quintana M et al., 2002, Curr Biol, Vol.12, p. 735-739. Also, “hsa-mir-125a” (miRBase Accession No. MI0000469,SEQ ID NO: 302) having a hairpin-like structure is known as a precursorof “hsa-miR-125a-3p”.

The term “hsa-miR-2861 gene” or “hsa-miR-2861” used herein includes thehsa-miR-2861 gene (miRBase Accession No. MIMAT0013802) described in SEQID NO: 110, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-2861 gene can be obtained by a methoddescribed in Li H et al., 2009, J Clin Invest, Vol. 119, p. 3666-3677.Also, “hsa-mir-2861” (miRBase Accession No. MI0013006, SEQ ID NO: 303)having a hairpin-like structure is known as a precursor of“hsa-miR-2861”.

The term “hsa-miR-6088 gene” or “hsa-miR-6088” used herein includes thehsa-miR-6088 gene (miRBase Accession No. MIMAT0023713) described in SEQID NO: 111, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-6088 gene can be obtained by a methoddescribed in Yoo J K et al., 2012, Stem Cells Dev, Vol. 21, p.2049-2057. Also, “hsa-mir-6088” (miRBase Accession No. MI0020365, SEQ IDNO: 304) having a hairpin-like structure is known as a precursor of“hsa-miR-6088”.

The term “hsa-miR-4758-5p gene” or “hsa-miR-4758-5p” used hereinincludes the hsa-miR-4758-5p gene (miRBase Accession No. MIMAT0019903)described in SEQ ID NO: 112, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-4758-5p gene can be obtainedby a method described in Persson H et al., 2011, Cancer Res, Vol. 71, p.78-86. Also, “hsa-mir-4758” (miRBase Accession No. MI0017399, SEQ ID NO:305) having a hairpin-like structure is known as a precursor of“hsa-miR-4758-5p”.

The term “hsa-miR-296-3p gene” or “hsa-miR-296-3p” used herein includesthe hsa-miR-296-3p gene (miRBase Accession No. MIMAT0004679) describedin SEQ ID NO: 113, a homolog or an ortholog of a different organismspecies, and the like. The hsa-miR-296-3p gene can be obtained by amethod described in Houbaviy H B et al., 2003, Dev Cell, Vol. 5, p.351-358. Also, “hsa-mir-296” (miRBase Accession No. MI0000747, SEQ IDNO: 306) having a hairpin-like structure is known as a precursor of“hsa-miR-296-3p”.

The term “hsa-miR-6738-5p gene” or “hsa-miR-6738-5p” used hereinincludes the hsa-miR-6738-5p gene (miRBase Accession No. MIMAT0027377)described in SEQ ID NO: 114, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6738-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6738” (miRBase Accession No. MI0022583, SEQ IDNO: 307) having a hairpin-like structure is known as a precursor of“hsa-miR-6738-5p”.

The term “hsa-miR-671-5p gene” or “hsa-miR-671-5p” used herein includesthe hsa-miR-671-5p gene (miRBase Accession No. MIMAT0003880) describedin SEQ ID NO: 115, a homolog or an ortholog of a different organismspecies, and the like. The hsa-miR-671-5p gene can be obtained by amethod described in Berezikov E et al., 2006, Genome Res, Vol. 16, p.1289-1298. Also, “hsa-mir-671” (miRBase Accession No. MI0003760, SEQ IDNO: 308) having a hairpin-like structure is known as a precursor of“hsa-miR-671-5p”.

The term “hsa-miR-4454 gene” or “hsa-miR-4454” used herein includes thehsa-miR-4454 gene (miRBase Accession No. MIMAT0018976) described in SEQID NO: 116, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-4454 gene can be obtained by a methoddescribed in Jima D D et al., 2010, Blood, Vol. 116, e118-e127. Also,“hsa-mir-4454” (miRBase Accession No. MI0016800, SEQ ID NO: 309) havinga hairpin-like structure is known as a precursor of “hsa-miR-4454”.

The term “hsa-miR-4516 gene” or “hsa-miR-4516” used herein includes thehsa-miR-4516 gene (miRBase Accession No. MIMAT0019053) described in SEQID NO: 117, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-4516 gene can be obtained by a methoddescribed in Jima D D et al., 2010, Blood, Vol. 116, e118-e127. Also,“hsa-mir-4516” (miRBase Accession No. MI0016882, SEQ ID NO: 310) havinga hairpin-like structure is known as a precursor of “hsa-miR-4516”.

The term “hsa-miR-7845-5p gene” or “hsa-miR-7845-5p” used hereinincludes the hsa-miR-7845-5p gene (miRBase Accession No. MIMAT0030420)described in SEQ ID NO: 118, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-7845-5p gene can be obtainedby a method described in Ple H et al., 2012, PLoS One, Vol. 7, e50746.Also, “hsa-mir-7845” (miRBase Accession No. MI0025515, SEQ ID NO: 311)having a hairpin-like structure is known as a precursor of“hsa-miR-7845-5p”.

The term “hsa-miR-4741 gene” or “hsa-miR-4741” used herein includes thehsa-miR-4741 gene (miRBase Accession No. MIMAT0019871) described in SEQID NO: 119, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-4741 gene can be obtained by a methoddescribed in Persson H et al., 2011, Cancer Res, Vol. 71, p. 78-86.Also, “hsa-mir-4741” (miRBase Accession No. MI0017379, SEQ ID NO: 312)having a hairpin-like structure is known as a precursor of“hsa-miR-4741”.

The term “hsa-miR-92b-5p gene” or “hsa-miR-92b-5p” used herein includesthe hsa-miR-92b-5p gene (miRBase Accession No. MIMAT0004792) describedin SEQ ID NO: 120, a homolog or an ortholog of a different organismspecies, and the like. The hsa-miR-92b-5p gene can be obtained by amethod described in Cummins J M et al., 2006, Proc Natl Acad Sci USA,Vol. 103, p. 3687-3692. Also, “hsa-mir-92b” (miRBase Accession No.MI0003560, SEQ ID NO: 313) having a hairpin-like structure is known as aprecursor of “hsa-miR-92b-5p”.

The term “hsa-miR-6795-5p gene” or “hsa-miR-6795-5p” used hereinincludes the hsa-miR-6795-5p gene (miRBase Accession No. MIMAT0027490)described in SEQ ID NO: 121, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6795-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6795” (miRBase Accession No. MI0022640, SEQ IDNO: 314) having a hairpin-like structure is known as a precursor of“hsa-miR-6795-5p”.

The term “hsa-miR-6805-3p gene” or “hsa-miR-6805-3p” used hereinincludes the hsa-miR-6805-3p gene (miRBase Accession No. MIMAT0027511)described in SEQ ID NO: 122, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6805-3p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6805” (miRBase Accession No. MI0022650, SEQ IDNO: 315) having a hairpin-like structure is known as a precursor of“hsa-miR-6805-3p”.

The term “hsa-miR-4725-3p gene” or “hsa-miR-4725-3p” used hereinincludes the hsa-miR-4725-3p gene (miRBase Accession No. MIMAT0019844)described in SEQ ID NO: 123, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-4725-3p gene can be obtainedby a method described in Persson H et al., 2011, Cancer Res, Vol. 71, p.78-86. Also, “hsa-mir-4725” (miRBase Accession No. MI0017362, SEQ ID NO:316) having a hairpin-like structure is known as a precursor of“hsa-miR-4725-3p”.

The term “hsa-miR-6782-5p gene” or “hsa-miR-6782-5p” used hereinincludes the hsa-miR-6782-5p gene (miRBase Accession No. MIMAT0027464)described in SEQ ID NO: 124, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6782-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6782” (miRBase Accession No. MI0022627, SEQ IDNO: 317) having a hairpin-like structure is known as a precursor of“hsa-miR-6782-5p”.

The term “hsa-miR-4688 gene” or “hsa-miR-4688” used herein includes thehsa-miR-4688 gene (miRBase Accession No. MIMAT0019777) described in SEQID NO: 125, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-4688 gene can be obtained by a methoddescribed in Persson H et al., 2011, Cancer Res, Vol. 71, p. 78-86.Also, “hsa-mir-4688” (miRBase Accession No. MI0017321, SEQ ID NO: 318)having a hairpin-like structure is known as a precursor of“hsa-miR-4688”.

The term “hsa-miR-6850-5p gene” or “hsa-miR-6850-5p” used hereinincludes the hsa-miR-6850-5p gene (miRBase Accession No. MIMAT0027600)described in SEQ ID NO: 126, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6850-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6850” (miRBase Accession No. MI0022696, SEQ IDNO: 319) having a hairpin-like structure is known as a precursor of“hsa-miR-6850-5p”.

The term “hsa-miR-6777-5p gene” or “hsa-miR-6777-5p” used hereinincludes the hsa-miR-6777-5p gene (miRBase Accession No. MIMAT0027454)described in SEQ ID NO: 127, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6777-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6777” (miRBase Accession No. MI0022622, SEQ IDNO: 320) having a hairpin-like structure is known as a precursor of“hsa-miR-6777-5p”.

The term “hsa-miR-6785-5p gene” or “hsa-miR-6785-5p” used hereinincludes the hsa-miR-6785-5p gene (miRBase Accession No. MIMAT0027470)described in SEQ ID NO: 128, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6785-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6785” (miRBase Accession No. MI0022630, SEQ IDNO: 321) having a hairpin-like structure is known as a precursor of“hsa-miR-6785-5p”.

The term “hsa-miR-7106-5p gene” or “hsa-miR-7106-5p” used hereinincludes the hsa-miR-7106-5p gene (miRBase Accession No. MIMAT0028109)described in SEQ ID NO: 129, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-7106-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-7106” (miRBase Accession No. MI0022957, SEQ IDNO: 322) having a hairpin-like structure is known as a precursor of“hsa-miR-7106-5p”.

The term “hsa-miR-3663-3p gene” or “hsa-miR-3663-3p” used hereinincludes the hsa-miR-3663-3p gene (miRBase Accession No. MIMAT0018085)described in SEQ ID NO: 130, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-3663-3p gene can be obtainedby a method described in Liao J Y et al., 2010, PLoS One, Vol. 5,e10563. Also, “hsa-mir-3663” (miRBase Accession No. MI0016064, SEQ IDNO: 323) having a hairpin-like structure is known as a precursor of“hsa-miR-3663-3p”.

The term “hsa-miR-6131 gene” or “hsa-miR-6131” used herein includes thehsa-miR-6131 gene (miRBase Accession No. MIMAT0024615) described in SEQID NO: 131, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-6131 gene can be obtained by a methoddescribed in Dannemann M et al., 2012, Genome Biol Evol, Vol. 4, p.552-564. Also, “hsa-mir-6131” (miRBase Accession No. MI0021276, SEQ IDNO: 324) having a hairpin-like structure is known as a precursor of“hsa-miR-6131”.

The term “hsa-miR-1915-3p gene” or “hsa-miR-1915-3p” used hereinincludes the hsa-miR-1915-3p gene (miRBase Accession No. MIMAT0007892)described in SEQ ID NO: 132, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-1915-3p gene can be obtainedby a method described in Bar M et al., 2008, Stem Cells, Vol. 26, p.2496-2505. Also, “hsa-mir-1915” (miRBase Accession No. MI0008336, SEQ IDNO: 288) having a hairpin-like structure is known as a precursor of“hsa-miR-1915-3p”.

The term “hsa-miR-4532 gene” or “hsa-miR-4532” used herein includes thehsa-miR-4532 gene (miRBase Accession No. MIMAT0019071) described in SEQID NO: 133, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-4532 gene can be obtained by a methoddescribed in Jima D D et al., 2010, Blood, Vol. 116, e118-e127. Also,“hsa-mir-4532” (miRBase Accession No. MI0016899, SEQ ID NO: 325) havinga hairpin-like structure is known as a precursor of “hsa-miR-4532”.

The term “hsa-miR-6820-5p gene” or “hsa-miR-6820-5p” used hereinincludes the hsa-miR-6820-5p gene (miRBase Accession No. MIMAT0027540)described in SEQ ID NO: 134, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6820-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6820” (miRBase Accession No. MI0022665, SEQ IDNO: 326) having a hairpin-like structure is known as a precursor of“hsa-miR-6820-5p”.

The term “hsa-miR-4689 gene” or “hsa-miR-4689” used herein includes thehsa-miR-4689 gene (miRBase Accession No. MIMAT0019778) described in SEQID NO: 135, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-4689 gene can be obtained by a methoddescribed in Persson H et al., 2011, Cancer Res, Vol. 71, p. 78-86.Also, “hsa-mir-4689” (miRBase Accession No. MI0017322, SEQ ID NO: 327)having a hairpin-like structure is known as a precursor of“hsa-miR-4689”.

The term “hsa-miR-4638-5p gene” or “hsa-miR-4638-5p” used hereinincludes the hsa-miR-4638-5p gene (miRBase Accession No. MIMAT0019695)described in SEQ ID NO: 136, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-4638-5p gene can be obtainedby a method described in Persson H et al., 2011, Cancer Res, Vol. 71, p.78-86. Also, “hsa-mir-4638” (miRBase Accession No. MI0017265, SEQ ID NO:328) having a hairpin-like structure is known as a precursor of“hsa-miR-4638-5p”.

The term “hsa-miR-3656 gene” or “hsa-miR-3656” used herein includes thehsa-miR-3656 gene (miRBase Accession No. MIMAT0018076) described in SEQID NO: 137, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-3656 gene can be obtained by a methoddescribed in Meiri E et al., 2010, Nucleic Acids Res, Vol. 38, p.6234-6246. Also, “hsa-mir-3656” (miRBase Accession No. MI0016056, SEQ IDNO: 329) having a hairpin-like structure is known as a precursor of“hsa-miR-3656”.

The term “hsa-miR-3621 gene” or “hsa-miR-3621” used herein includes thehsa-miR-3621 gene (miRBase Accession No. MIMAT0018002) described in SEQID NO: 138, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-3621 gene can be obtained by a methoddescribed in Witten D et al., 2010, BMC Biol, Vol. 8, p. 58. Also,“hsa-mir-3621” (miRBase Accession No. MI0016012, SEQ ID NO: 330) havinga hairpin-like structure is known as a precursor of “hsa-miR-3621”.

The term “hsa-miR-6769b-5p gene” or “hsa-miR-6769b-5p” used hereinincludes the hsa-miR-6769b-5p gene (miRBase Accession No. MIMAT0027620)described in SEQ ID NO: 139, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6769b-5p gene can beobtained by a method described in Ladewig E et al., 2012, Genome Res,Vol. 22, p. 1634-1645. Also, “hsa-mir-6769b” (miRBase Accession No.MI0022706, SEQ ID NO: 331) having a hairpin-like structure is known as aprecursor of “hsa-miR-6769b-5p”.

The term “hsa-miR-149-3p gene” or “hsa-miR-149-3p” used herein includesthe hsa-miR-149-3p gene (miRBase Accession No. MIMAT0004609) describedin SEQ ID NO: 140, a homolog or an ortholog of a different organismspecies, and the like. The hsa-miR-149-3p gene can be obtained by amethod described in Lagos-Quintana M et al., 2002, Curr Biol, Vol. 12,p. 735-739. Also, “hsa-mir-149” (miRBase Accession No. MI0000478, SEQ IDNO: 332) having a hairpin-like structure is known as a precursor of“hsa-miR-149-3p”.

The term “hsa-miR-23b-3p gene” or “hsa-miR-23b-3p” used herein includesthe hsa-miR-23b-3p gene (miRBase Accession No. MIMAT0000418) describedin SEQ ID NO: 141, a homolog or an ortholog of a different organismspecies, and the like. The hsa-miR-23b-3p gene can be obtained by amethod described in Lagos-Quintana M et al., 2002, Curr Biol, Vol. 12,p. 735-739. Also, “hsa-mir-23b” (miRBase Accession No. MI0000439, SEQ IDNO: 333) having a hairpin-like structure is known as a precursor of“hsa-miR-23b-3p”.

The term “hsa-miR-3135b gene” or “hsa-miR-3135b” used herein includesthe hsa-miR-3135b gene (miRBase Accession No. MIMAT0018985) described inSEQ ID NO: 142, a homolog or an ortholog of a different organismspecies, and the like. The hsa-miR-3135b gene can be obtained by amethod described in Jima D D et al., 2010, Blood, Vol. 116, e118-e127.Also, “hsa-mir-3135b” (miRBase Accession No. MI0016809, SEQ ID NO: 334)having a hairpin-like structure is known as a precursor of“hsa-miR-3135b”.

The term “hsa-miR-6848-5p gene” or “hsa-miR-6848-5p” used hereinincludes the hsa-miR-6848-5p gene (miRBase Accession No. MIMAT0027596)described in SEQ ID NO: 143, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6848-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6848” (miRBase Accession No. MI0022694, SEQ IDNO: 335) having a hairpin-like structure is known as a precursor of“hsa-miR-6848-5p”.

The term “hsa-miR-6769a-5p gene” or “hsa-miR-6769a-5p” used hereinincludes the hsa-miR-6769a-5p gene (miRBase Accession No. MIMAT0027438)described in SEQ ID NO: 144, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6769a-5p gene can beobtained by a method described in Ladewig E et al., 2012, Genome Res,Vol. 22, p. 1634-1645. Also, “hsa-mir-6769a” (miRBase Accession No.MI0022614, SEQ ID NO: 336) having a hairpin-like structure is known as aprecursor of “hsa-miR-6769a-5p”.

The term “hsa-miR-4327 gene” or “hsa-miR-4327” used herein includes thehsa-miR-4327 gene (miRBase Accession No. MIMAT0016889) described in SEQID NO: 145, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-4327 gene can be obtained by a methoddescribed in Goff L A et al., 2009, PLoS One, Vol. 4, e7192. Also,“hsa-mir-4327” (miRBase Accession No. MI0015867, SEQ ID NO: 337) havinga hairpin-like structure is known as a precursor of “hsa-miR-4327”.

The term “hsa-miR-6765-3p gene” or “hsa-miR-6765-3p” used hereinincludes the hsa-miR-6765-3p gene (miRBase Accession No. MIMAT0027431)described in SEQ ID NO: 146, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6765-3p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6765” (miRBase Accession No. MI0022610, SEQ IDNO: 338) having a hairpin-like structure is known as a precursor of“hsa-miR-6765-3p”.

The term “hsa-miR-6716-5p gene” or “hsa-miR-6716-5p” used hereinincludes the hsa-miR-6716-5p gene (miRBase Accession No. MIMAT0025844)described in SEQ ID NO: 147, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6716-5p gene can be obtainedby a method described in Li Y et al., 2012, Gene, Vol. 497, p. 330-335.Also, “hsa-mir-6716” (miRBase Accession No. MI0022550, SEQ ID NO: 339)having a hairpin-like structure is known as a precursor of“hsa-miR-6716-5p”.

The term “hsa-miR-6877-5p gene” or “hsa-miR-6877-5p” used hereinincludes the hsa-miR-6877-5p gene (miRBase Accession No. MIMAT0027654)described in SEQ ID NO: 148, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6877-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6877” (miRBase Accession No. MI0022724, SEQ IDNO: 340) having a hairpin-like structure is known as a precursor of“hsa-miR-6877-5p”.

The term “hsa-miR-6727-5p gene” or “hsa-miR-6727-5p” used hereinincludes the hsa-miR-6727-5p gene (miRBase Accession No. MIMAT0027355)described in SEQ ID NO: 149, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6727-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6727” (miRBase Accession No. MI0022572, SEQ IDNO: 341) having a hairpin-like structure is known as a precursor of“hsa-miR-6727-5p”.

The term “hsa-miR-4534 gene” or “hsa-miR-4534” used herein includes thehsa-miR-4534 gene (miRBase Accession No. MIMAT0019073) described in SEQID NO: 150, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-4534 gene can be obtained by a methoddescribed in Jima D D et al., 2010, Blood, Vol. 116, e118-e127. Also,“hsa-mir-4534” (miRBase Accession No. MI0016901, SEQ ID NO: 342) havinga hairpin-like structure is known as a precursor of “hsa-miR-4534”.

The term “hsa-miR-614 gene” or “hsa-miR-614” used herein includes thehsa-miR-614 gene (miRBase Accession No. MIMAT0003282) described in SEQID NO: 151, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-614 gene can be obtained by a method describedin Cummins J M et al., 2006, Proc Natl Acad Sci USA, Vol. 103, p.3687-3692. Also, “hsa-mir-614” (miRBase Accession No. MI0003627, SEQ IDNO: 343) having a hairpin-like structure is known as a precursor of“hsa-miR-614”.

The term “hsa-miR-1202 gene” or “hsa-miR-1202” used herein includes thehsa-miR-1202 gene (miRBase Accession No. MIMAT0005865) described in SEQID NO: 152, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-1202 gene can be obtained by a methoddescribed in Marton S et al., 2008, Leukemia, Vol. 22, p. 330-338. Also,“hsa-mir-1202” (miRBase Accession No. MI0006334, SEQ ID NO: 344) havinga hairpin-like structure is known as a precursor of “hsa-miR-1202”.

The term “hsa-miR-575 gene” or “hsa-miR-575” used herein includes thehsa-miR-575 gene (miRBase Accession No. MIMAT0003240) described in SEQID NO: 153, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-575 gene can be obtained by a method describedin Cummins J M et al., 2006, Proc Natl Acad Sci USA, Vol. 103, p.3687-3692. Also, “hsa-mir-575” (miRBase Accession No. MI0003582, SEQ IDNO: 345) having a hairpin-like structure is known as a precursor of“hsa-miR-575”.

The term “hsa-miR-6870-5p gene” or “hsa-miR-6870-5p” used hereinincludes the hsa-miR-6870-5p gene (miRBase Accession No. MIMAT0027640)described in SEQ ID NO: 154, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6870-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6870” (miRBase Accession No. MI0022717, SEQ IDNO: 346) having a hairpin-like structure is known as a precursor of“hsa-miR-6870-5p”.

The term “hsa-miR-6722-3p gene” or “hsa-miR-6722-3p” used hereinincludes the hsa-miR-6722-3p gene (miRBase Accession No. MIMAT0025854)described in SEQ ID NO: 155, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6722-3p gene can be obtainedby a method described in Li Y et al., 2012, Gene, Vol. 497, p. 330-335.Also, “hsa-mir-6722” (miRBase Accession No. MI0022557, SEQ ID NO: 347)having a hairpin-like structure is known as a precursor of“hsa-miR-6722-3p”.

The term “hsa-miR-7977 gene” or “hsa-miR-7977” used herein includes thehsa-miR-7977 gene (miRBase Accession No. MIMAT0031180) described in SEQID NO: 156, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-7977 gene can be obtained by a methoddescribed in Velthut-Meikas A et al., 2013, Mol Endocrinol, online.Also, “hsa-mir-7977” (miRBase Accession No. MI0025753, SEQ ID NO: 348)having a hairpin-like structure is known as a precursor of“hsa-miR-7977”.

The term “hsa-miR-4649-5p gene” or “hsa-miR-4649-5p” used hereinincludes the hsa-miR-4649-5p gene (miRBase Accession No. MIMAT0019711)described in SEQ ID NO: 157, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-4649-5p gene can be obtainedby a method described in Persson H et al., 2011, Cancer Res, Vol. 71, p.78-86. Also, “hsa-mir-4649” (miRBase Accession No. MI0017276, SEQ ID NO:349) having a hairpin-like structure is known as a precursor of“hsa-miR-4649-5p”.

The term “hsa-miR-4675 gene” or “hsa-miR-4675” used herein includes thehsa-miR-4675 gene (miRBase Accession No. MIMAT0019757) described in SEQID NO: 158, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-4675 gene can be obtained by a methoddescribed in Persson H et al., 2011, Cancer Res, Vol. 71, p. 78-86.Also, “hsa-mir-4675” (miRBase Accession No. MI0017306, SEQ ID NO: 350)having a hairpin-like structure is known as a precursor of“hsa-miR-4675”.

The term “hsa-miR-6075 gene” or “hsa-miR-6075” used herein includes thehsa-miR-6075 gene (miRBase Accession No. MIMAT0023700) described in SEQID NO: 159, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-6075 gene can be obtained by a methoddescribed in Voellenkle C et al., 2012, RNA, Vol. 18, p. 472-484. Also,“hsa-mir-6075” (miRBase Accession No. MI0020352, SEQ ID NO: 351) havinga hairpin-like structure is known as a precursor of “hsa-miR-6075”.

The term “hsa-miR-6779-5p gene” or “hsa-miR-6779-5p” used hereinincludes the hsa-miR-6779-5p gene (miRBase Accession No. MIMAT0027458)described in SEQ ID NO: 160, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6779-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6779” (miRBase Accession No. MI0022624, SEQ IDNO: 352) having a hairpin-like structure is known as a precursor of“hsa-miR-6779-5p”.

The term “hsa-miR-4271 gene” or “hsa-miR-4271” used herein includes thehsa-miR-4271 gene (miRBase Accession No. MIMAT0016901) described in SEQID NO: 161, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-4271 gene can be obtained by a methoddescribed in Goff L A et al., 2009, PLoS One, Vol. 4, e7192. Also,“hsa-mir-4271” (miRBase Accession No. MI0015879, SEQ ID NO: 353) havinga hairpin-like structure is known as a precursor of “hsa-miR-4271”.

The term “hsa-miR-3196 gene” or “hsa-miR-3196” used herein includes thehsa-miR-3196 gene (miRBase Accession No. MIMAT0015080) described in SEQID NO: 162, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-3196 gene can be obtained by a methoddescribed in Stark M S et al., 2010, PLoS One, Vol. 5, e9685. Also,“hsa-mir-3196” (miRBase Accession No. MI0014241, SEQ ID NO: 354) havinga hairpin-like structure is known as a precursor of “hsa-miR-3196”.

The term “hsa-miR-6803-5p gene” or “hsa-miR-6803-5p” used hereinincludes the hsa-miR-6803-5p gene (miRBase Accession No. MIMAT0027506)described in SEQ ID NO: 163, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6803-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6803” (miRBase Accession No. MI0022648, SEQ IDNO: 355) having a hairpin-like structure is known as a precursor of“hsa-miR-6803-5p”.

The term “hsa-miR-6789-5p gene” or “hsa-miR-6789-5p” used hereinincludes the hsa-miR-6789-5p gene (miRBase Accession No. MIMAT0027478)described in SEQ ID NO: 164, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6789-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6789” (miRBase Accession No. MI0022634, SEQ IDNO: 356) having a hairpin-like structure is known as a precursor of“hsa-miR-6789-5p”.

The term “hsa-miR-4648 gene” or “hsa-miR-4648” used herein includes thehsa-miR-4648 gene (miRBase Accession No. MIMAT0019710) described in SEQID NO: 165, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-4648 gene can be obtained by a methoddescribed in Persson H et al., 2011, Cancer Res, Vol. 71, p. 78-86.Also, “hsa-mir-4648” (miRBase Accession No. MI0017275, SEQ ID NO: 357)having a hairpin-like structure is known as a precursor of“hsa-miR-4648”.

The term “hsa-miR-4508 gene” or “hsa-miR-4508” used herein includes thehsa-miR-4508 gene (miRBase Accession No. MIMAT0019045) described in SEQID NO: 166, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-4508 gene can be obtained by a methoddescribed in Jima D D et al., 2010, Blood, Vol. 116, e118-e127. Also,“hsa-mir-4508” (miRBase Accession No. MI0016872, SEQ ID NO: 358) havinga hairpin-like structure is known as a precursor of “hsa-miR-4508”.

The term “hsa-miR-4749-5p gene” or “hsa-miR-4749-5p” used hereinincludes the hsa-miR-4749-5p gene (miRBase Accession No. MIMAT0019885)described in SEQ ID NO: 167, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-4749-5p gene can be obtainedby a method described in Persson H et al., 2011, Cancer Res, Vol. 71, p.78-86. Also, “hsa-mir-4749” (miRBase Accession No. MI0017388, SEQ ID NO:359) having a hairpin-like structure is known as a precursor of“hsa-miR-4749-5p”.

The term “hsa-miR-4505 gene” or “hsa-miR-4505” used herein includes thehsa-miR-4505 gene (miRBase Accession No. MIMAT0019041) described in SEQID NO: 168, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-4505 gene can be obtained by a methoddescribed in Jima D D et al., 2010, Blood, Vol. 116, e118-e127. Also,“hsa-mir-4505” (miRBase Accession No. MI0016868, SEQ ID NO: 360) havinga hairpin-like structure is known as a precursor of “hsa-miR-4505”.

The term “hsa-miR-5698 gene” or “hsa-miR-5698” used herein includes thehsa-miR-5698 gene (miRBase Accession No. MIMAT0022491) described in SEQID NO: 169, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-5698 gene can be obtained by a methoddescribed in Watahiki A et al., 2011, PLoS One, Vol. 6, e24950. Also,“hsa-mir-5698” (miRBase Accession No. MI0019305, SEQ ID NO: 361) havinga hairpin-like structure is known as a precursor of “hsa-miR-5698”.

The term “hsa-miR-1199-5p gene” or “hsa-miR-1199-5p” used hereinincludes the hsa-miR-1199-5p gene (miRBase Accession No. MIMAT0031119)described in SEQ ID NO: 170, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-1199-5p gene can be obtainedby a method described in Salvi A et al., 2013, Int J Oncol, Vol. 42, p.391-402. Also, “hsa-mir-1199” (miRBase Accession No. MI0020340, SEQ IDNO: 362) having a hairpin-like structure is known as a precursor of“hsa-miR-1199-5p”.

The term “hsa-miR-4763-3p gene” or “hsa-miR-4763-3p” used hereinincludes the hsa-miR-4763-3p gene (miRBase Accession No. MIMAT0019913)described in SEQ ID NO: 171, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-4763-3p gene can be obtainedby a method described in Persson H et al., 2011, Cancer Res, Vol. 71, p.78-86. Also, “hsa-mir-4763” (miRBase Accession No. MI0017404, SEQ ID NO:363) having a hairpin-like structure is known as a precursor of“hsa-miR-4763-3p”.

The term “hsa-miR-1231 gene” or “hsa-miR-1231” used herein includes thehsa-miR-1231 gene (miRBase Accession No. MIMAT0005586) described in SEQID NO: 172, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-1231 gene can be obtained by a methoddescribed in Berezikov E et al., 2007, Mol Cell, Vol. 28, p. 328-336.Also, “hsa-mir-1231” (miRBase Accession No. MI0006321, SEQ ID NO: 364)having a hairpin-like structure is known as a precursor of“hsa-miR-1231”.

The term “hsa-miR-1233-5p gene” or “hsa-miR-1233-5p” used hereinincludes the hsa-miR-1233-5p gene (miRBase Accession No. MIMAT0022943)described in SEQ ID NO: 173, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-1233-5p gene can be obtainedby a method described in Berezikov E et al., 2007, Mol Cell, Vol. 28, p.328-336. Also, “hsa-mir-1233-1 and hsa-mir-1233-2” (miRBase AccessionNos. MI0006323 and MI0015973, SEQ ID NOs: 365 and 366) having ahairpin-like structure are known as precursors of “hsa-miR-1233-5p”.

The term “hsa-miR-150-3p gene” or “hsa-miR-150-3p” used herein includesthe hsa-miR-150-3p gene (miRBase Accession No. MIMAT0004610) describedin SEQ ID NO: 174, a homolog or an ortholog of a different organismspecies, and the like. The hsa-miR-150-3p gene can be obtained by amethod described in Lagos-Quintana M et al., 2002, Curr Biol, Vol. 12,p. 735-739. Also, “hsa-mir-150” (miRBase Accession No. MI0000479, SEQ IDNO: 367) having a hairpin-like structure is known as a precursor of“hsa-miR-150-3p”.

The term “hsa-miR-1225-3p gene” or “hsa-miR-1225-3p” used hereinincludes the hsa-miR-1225-3p gene (miRBase Accession No. MIMAT0005573)described in SEQ ID NO: 175, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-1225-3p gene can be obtainedby a method described in Berezikov E et al., 2007, Mol Cell, Vol. 28, p.328-336. Also, “hsa-mir-1225” (miRBase Accession No. MI0006311, SEQ IDNO: 235) having a hairpin-like structure is known as a precursor of“hsa-miR-1225-3p”.

The term “hsa-miR-92a-2-5p gene” or “hsa-miR-92a-2-5p” used hereinincludes the hsa-miR-92a-2-5p gene (miRBase Accession No. MIMAT0004508)described in SEQ ID NO: 176, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-92a-2-5p gene can beobtained by a method described in Mourelatos Z et al., 2002, Genes Dev,Vol. 16, p. 720-728. Also, “hsa-mir-92a-2” (miRBase Accession No.MI0000094, SEQ ID NO: 368) having a hairpin-like structure is known as aprecursor of “hsa-miR-92a-2-5p”.

The term “hsa-miR-423-5p gene” or “hsa-miR-423-5p” used herein includesthe hsa-miR-423-5p gene (miRBase Accession No. MIMAT0004748) describedin SEQ ID NO: 177, a homolog or an ortholog of a different organismspecies, and the like. The hsa-miR-423-5p gene can be obtained by amethod described in Kasashima K et al., 2004, Biochem Biophys ResCommun, Vol. 322, p. 403-410. Also, “hsa-mir-423” (miRBase Accession No.MI0001445, SEQ ID NO: 369) having a hairpin-like structure is known as aprecursor of “hsa-miR-423-5p”.

The term “hsa-miR-1268a gene” or “hsa-miR-1268a” used herein includesthe hsa-miR-1268a gene (miRBase Accession No. MIMAT0005922) described inSEQ ID NO: 178, a homolog or an ortholog of a different organismspecies, and the like. The hsa-miR-1268a gene can be obtained by amethod described in Morin R D et al., 2008, Genome Res, Vol. 18, p.610-621. Also, “hsa-mir-1268a” (miRBase Accession No. MI0006405, SEQ IDNO: 370) having a hairpin-like structure is known as a precursor of“hsa-miR-1268a”.

The term “hsa-miR-128-2-5p gene” or “hsa-miR-128-2-5p” used hereinincludes the hsa-miR-128-2-5p gene (miRBase Accession No. MIMAT0031095)described in SEQ ID NO: 179, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-128-2-5p gene can beobtained by a method described in Lagos-Quintana M et al., 2002, CurrBiol, Vol. 12, p. 735-739. Also, “hsa-mir-128-2” (miRBase Accession No.MI0000727, SEQ ID NO: 371) having a hairpin-like structure is known as aprecursor of “hsa-miR-128-2-5p”.

The term “hsa-miR-24-3p gene” or “hsa-miR-24-3p” used herein includesthe hsa-miR-24-3p gene (miRBase Accession No. MIMAT0000080) described inSEQ ID NO: 180, a homolog or an ortholog of a different organismspecies, and the like. The hsa-miR-24-3p gene can be obtained by amethod described in Lagos-Quintana M et al., 2001, Science, Vol. 294, p.853-858. Also, “hsa-mir-24-1 and hsa-mir-24-2” (miRBase Accession Nos.MI0000080 and MI0000081, SEQ ID NOs: 372 and 373) having a hairpin-likestructure are known as precursors of “hsa-miR-24-3p”.

The term “hsa-miR-4697-5p gene” or “hsa-miR-4697-5p” used hereinincludes the hsa-miR-4697-5p gene (miRBase Accession No. MIMAT0019791)described in SEQ ID NO: 181, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-4697-5p gene can be obtainedby a method described in Persson H et al., 2011, Cancer Res, Vol. 71, p.78-86. Also, “hsa-mir-4697” (miRBase Accession No. MI0017330, SEQ ID NO:374) having a hairpin-like structure is known as a precursor of“hsa-miR-4697-5p”.

The term “hsa-miR-3197 gene” or “hsa-miR-3197” used herein includes thehsa-miR-3197 gene (miRBase Accession No. MIMAT0015082) described in SEQID NO: 182, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-3197 gene can be obtained by a methoddescribed in Stark M S et al., 2010, PLoS One, Vol. 5, e9685. Also,“hsa-mir-3197” (miRBase Accession No. MI0014245, SEQ ID NO: 375) havinga hairpin-like structure is known as a precursor of “hsa-miR-3197”.

The term “hsa-miR-675-5p gene” or “hsa-miR-675-5p” used herein includesthe hsa-miR-675-5p gene (miRBase Accession No. MIMAT0004284) describedin SEQ ID NO: 183, a homolog or an ortholog of a different organismspecies, and the like. The hsa-miR-675-5p gene can be obtained by amethod described in Cai X et al., 2007, RNA, Vol. 13, p. 313-316. Also,“hsa-mir-675” (miRBase Accession No. MI0005416, SEQ ID NO: 376) having ahairpin-like structure is known as a precursor of “hsa-miR-675-5p”.

The term “hsa-miR-4486 gene” or “hsa-miR-4486” used herein includes thehsa-miR-4486 gene (miRBase Accession No. MIMAT0019020) described in SEQID NO: 184, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-4486 gene can be obtained by a methoddescribed in Jima D D et al., 2010, Blood, Vol. 116, e118-e127. Also,“hsa-mir-4486” (miRBase Accession No. MI0016847, SEQ ID NO: 377) havinga hairpin-like structure is known as a precursor of “hsa-miR-4486”.

The term “hsa-miR-7107-5p gene” or “hsa-miR-7107-5p” used hereinincludes the hsa-miR-7107-5p gene (miRBase Accession No. MIMAT0028111)described in SEQ ID NO: 185, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-7107-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-7107” (miRBase Accession No. MI0022958, SEQ IDNO: 378) having a hairpin-like structure is known as a precursor of“hsa-miR-7107-5p”.

The term “hsa-miR-23a-3p gene” or “hsa-miR-23a-3p” used herein includesthe hsa-miR-23a-3p gene (miRBase Accession No. MIMAT0000078) describedin SEQ ID NO: 186, a homolog or an ortholog of a different organismspecies, and the like. The hsa-miR-23a-3p gene can be obtained by amethod described in Lagos-Quintana M et al., 2001, Science, Vol. 294, p.853-858. Also, “hsa-mir-23a” (miRBase Accession No. MI0000079, SEQ IDNO: 379) having a hairpin-like structure is known as a precursor of“hsa-miR-23a-3p”.

The term “hsa-miR-4667-5p gene” or “hsa-miR-4667-5p” used hereinincludes the hsa-miR-4667-5p gene (miRBase Accession No. MIMAT0019743)described in SEQ ID NO: 187, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-4667-5p gene can be obtainedby a method described in Persson H et al., 2011, Cancer Res, Vol. 71, p.78-86. Also, “hsa-mir-4667” (miRBase Accession No. MI0017297, SEQ ID NO:380) having a hairpin-like structure is known as a precursor of“hsa-miR-4667-5p”.

The term “hsa-miR-451a gene” or “hsa-miR-451a” used herein includes thehsa-miR-451a gene (miRBase Accession No. MIMAT0001631) described in SEQID NO: 188, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-451a gene can be obtained by a methoddescribed in Altuvia Y et al., 2005, Nucleic Acids Res, Vol. 33, p.2697-2706. Also, “hsa-mir-451a” (miRBase Accession No. MI0001729, SEQ IDNO: 381) having a hairpin-like structure is known as a precursor of“hsa-miR-451a”.

The term “hsa-miR-3940-5p gene” or “hsa-miR-3940-5p” used hereinincludes the hsa-miR-3940-5p gene (miRBase Accession No. MIMAT0019229)described in SEQ ID NO: 189, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-3940-5p gene can be obtainedby a method described in Liao J Y et al., 2010, PLoS One, Vol. 5,e10563. Also, “hsa-mir-3940” (miRBase Accession No. MI0016597, SEQ IDNO: 382) having a hairpin-like structure is known as a precursor of“hsa-miR-3940-5p”.

The term “hsa-miR-8059 gene” or “hsa-miR-8059” used herein includes thehsa-miR-8059 gene (miRBase Accession No. MIMAT0030986) described in SEQID NO: 190, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-8059 gene can be obtained by a methoddescribed in Wang H J et al., 2013, Shock, Vol. 39, p. 480-487. Also,“hsa-mir-8059” (miRBase Accession No. MI0025895, SEQ ID NO: 383) havinga hairpin-like structure is known as a precursor of “hsa-miR-8059”.

The term “hsa-miR-6813-5p gene” or “hsa-miR-6813-5p” used hereinincludes the hsa-miR-6813-5p gene (miRBase Accession No. MIMAT0027526)described in SEQ ID NO: 191, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6813-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6813” (miRBase Accession No. MI0022658, SEQ IDNO: 384) having a hairpin-like structure is known as a precursor of“hsa-miR-6813-5p”.

The term “hsa-miR-4492 gene” or “hsa-miR-4492” used herein includes thehsa-miR-4492 gene (miRBase Accession No. MIMAT0019027) described in SEQID NO: 192, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-4492 gene can be obtained by a methoddescribed in Jima D D et al., 2010, Blood, Vol. 116, e118-e127. Also,“hsa-mir-4492” (miRBase Accession No. MI0016854, SEQ ID NO: 385) havinga hairpin-like structure is known as a precursor of “hsa-miR-4492”.

The term “hsa-miR-4476 gene” or “hsa-miR-4476” used herein includes thehsa-miR-4476 gene (miRBase Accession No. MIMAT0019003) described in SEQID NO: 193, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-4476 gene can be obtained by a methoddescribed in Jima D D et al., 2010, Blood, Vol. 116, e118-e127. Also,“hsa-mir-4476” (miRBase Accession No. MI0016828, SEQ ID NO: 386) havinga hairpin-like structure is known as a precursor of “hsa-miR-4476”.

The term “hsa-miR-6090 gene” or “hsa-miR-6090” used herein includes thehsa-miR-6090 gene (miRBase Accession No. MIMAT0023715) described in SEQID NO: 194, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-6090 gene can be obtained by a methoddescribed in Yoo J K et al., 2012, Stem Cells Dev, Vol. 21, p.2049-2057. Also, “hsa-mir-6090” (miRBase Accession No. MI0020367, SEQ IDNO: 387) having a hairpin-like structure is known as a precursor of“hsa-miR-6090”.

The term “hsa-miR-6836-3p gene” or “hsa-miR-6836-3p” used hereinincludes the hsa-miR-6836-3p gene (miRBase Accession No. MIMAT0027575)described in SEQ ID NO: 606, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6836-3p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6836” (miRBase Accession No. MI0022682, SEQ IDNO: 615) having a hairpin-like structure is known as a precursor of“hsa-miR-6836-3p”.

The term “hsa-miR-3195 gene” or “hsa-miR-3195” used herein includes thehsa-miR-3195 gene (miRBase Accession No. MIMAT0015079) described in SEQID NO: 607, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-3195 gene can be obtained by a methoddescribed in Stark M S et al., 2010, PLoS One, Vol. 5, e9685. Also,“hsa-mir-3195” (miRBase Accession No. MI0014240, SEQ ID NO: 616) havinga hairpin-like structure is known as a precursor of “hsa-miR-3195”.

The term “hsa-miR-718 gene” or “hsa-miR-718” used herein includes thehsa-miR-718 gene (miRBase Accession No. MIMAT0012735) described in SEQID NO: 608, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-718 gene can be obtained by a method describedin Artzi S et al., 2008, BMC Bioinformatics, Vol. 9, p. 39. Also,“hsa-mir-718” (miRBase Accession No. MI0012489, SEQ ID NO: 617) having ahairpin-like structure is known as a precursor of “hsa-miR-718”.

The term “hsa-miR-3178 gene” or “hsa-miR-3178” used herein includes thehsa-miR-3178 gene (miRBase Accession No. MIMAT0015055) described in SEQID NO: 609, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-3178 gene can be obtained by a methoddescribed in Stark M S et al., 2010, PLoS One, Vol. 5, e9685. Also,“hsa-mir-3178” (miRBase Accession No. MI0014212, SEQ ID NO: 618) havinga hairpin-like structure is known as a precursor of “hsa-miR-3178”.

The term “hsa-miR-638 gene” or “hsa-miR-638” used herein includes thehsa-miR-638 gene (miRBase Accession No. MIMAT0003308) described in SEQID NO: 610, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-638 gene can be obtained by a method describedin Cummins J M et al., 2006, Proc Natl Acad Sci USA, Vol. 103, p.3687-3692. Also, “hsa-mir-638” (miRBase Accession No. MI0003653, SEQ IDNO: 619) having a hairpin-like structure is known as a precursor of“hsa-miR-638”.

The term “hsa-miR-4497 gene” or “hsa-miR-4497” used herein includes thehsa-miR-4497 gene (miRBase Accession No. MIMAT0019032) described in SEQID NO: 611, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-4497 gene can be obtained by a methoddescribed in Jima D D et al., 2010, Blood, Vol. 116, e118-e127. Also,“hsa-mir-4497” (miRBase Accession No. MI0016859, SEQ ID NO: 620) havinga hairpin-like structure is known as a precursor of “hsa-miR-4497”.

The term “hsa-miR-6085 gene” or “hsa-miR-6085” used herein includes thehsa-miR-6085 gene (miRBase Accession No. MIMAT0023710) described in SEQID NO: 612, a homolog or an ortholog of a different organism species,and the like. The hsa-miR-6085 gene can be obtained by a methoddescribed in Voellenkle C et al., 2012, RNA, Vol. 18, p. 472-484. Also,“hsa-mir-6085” (miRBase Accession No. MI0020362, SEQ ID NO: 621) havinga hairpin-like structure is known as a precursor of “hsa-miR-6085”.

The term “hsa-miR-6752-5p gene” or “hsa-miR-6752-5p” used hereinincludes the hsa-miR-6752-5p gene (miRBase Accession No. MIMAT0027404)described in SEQ ID NO: 613, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-6752-5p gene can be obtainedby a method described in Ladewig E et al., 2012, Genome Res, Vol. 22, p.1634-1645. Also, “hsa-mir-6752” (miRBase Accession No. MI0022597, SEQ IDNO: 622) having a hairpin-like structure is known as a precursor of“hsa-miR-6752-5p”.

The term “hsa-miR-135a-3p gene” or “hsa-miR-135a-3p” used hereinincludes the hsa-miR-135a-3p gene (miRBase Accession No. MIMAT0004595)described in SEQ ID NO: 614, a homolog or an ortholog of a differentorganism species, and the like. The hsa-miR-135a-3p gene can be obtainedby a method described in Lagos-Quintana M et al., 2002, Curr Biol, Vol.12, p. 735-739. Also, “hsa-mir-135a” (miRBase Accession No. MI0000452,SEQ ID NO: 623) having a hairpin-like structure is known as a precursorof “hsa-miR-135a-3p”.

A mature miRNA may become a variant due to the sequence that is cleavedshorter or longer by one to several upstream or downstream nucleotidesor nucleotide substitution when cut out as the mature miRNA from its RNAprecursor having a hairpin-like structure. This variant is called isomiR(Morin R D. et al., 2008, Genome Res., Vol. 18, p. 610-621). The miRBaseRelease 20 shows the nucleotide sequences represented by SEQ ID NOs: 1to 194 and 606 to 614 as well as a large number of the nucleotidesequence variants and fragments represented by SEQ ID NOs: 388 to 605and 624 to 635, which are called isomiRs. These variants can also beobtained as miRNAs having a nucleotide sequence represented by any ofSEQ ID NOs: 1 to 194 and 606 to 614. Specifically, among the variants ofpolynucleotides consisting of the nucleotide sequence represented by anyof SEQ ID NOs: 5, 7, 8, 9, 11, 16, 19, 20, 21, 26, 27, 28, 30, 34, 37,38, 39, 41, 43, 45, 46, 48, 50, 54, 55, 57, 58, 61, 62, 63, 64, 65, 66,67, 69, 70, 71, 73, 74, 76, 77, 78, 80, 81, 82, 84, 85, 86, 88, 89, 94,95, 97, 98, 99, 100, 101, 104, 107, 108, 109, 110, 111, 112, 113, 115,116, 117, 119, 120, 123, 125, 131, 132, 133, 135, 136, 137, 140, 141,142, 147, 151, 152, 157, 161, 162, 165, 166, 167, 168, 169, 171, 173,174, 176, 177, 178, 179, 180, 182, 183, 184, 186, 187, 188, 189, 192,193, 607, 608, 609, 610, 611 and 614, or a nucleotide sequence derivedfrom the nucleotide sequence by the replacement of u with t according tothe present invention, examples of the longest variants registered inthe miRBase Release 20 include polynucleotides represented by SEQ IDNOs:388, 390, 392, 394, 396, 398, 400, 402, 404, 406, 408, 410, 412,414, 416, 418, 420, 422, 424, 426, 428, 430, 432, 434, 436, 438, 440,442, 444, 446, 448, 450, 452, 454, 456, 458, 460, 462, 464, 466, 468,470, 472, 474, 476, 478, 480, 482, 484, 486, 488, 490, 492, 494, 496,498, 500, 502, 504, 506, 508, 510, 512, 514, 516, 518, 520, 522, 524,526, 528, 530, 532, 534, 536, 538, 540, 542, 544, 546, 548, 550, 552,554, 556, 558, 560, 562, 564, 566, 568, 570, 572, 574, 576, 578, 580,582, 584, 586, 588, 590, 592, 594, 596, 598, 600, 602, 604, 624, 626,628, 630, 632 and 634, respectively. Also, among the variants ofpolynucleotides consisting of a nucleotide sequence represented by anyof SEQ ID NOs: 5, 7, 8, 9, 11, 16, 19, 20, 21, 26, 27, 28, 30, 34, 37,38, 39, 41, 43, 45, 46, 48, 50, 54, 55, 57, 58, 61, 62, 63, 64, 65, 66,67, 69, 70, 71, 73, 74, 76, 77, 78, 80, 81, 82, 84, 85, 86, 88, 89, 94,95, 97, 98, 99, 100, 101, 104, 107, 108, 109, 110, 111, 112, 113, 115,116, 117, 119, 120, 123, 125, 131, 132, 133, 135, 136, 137, 140, 141,142, 147, 151, 152, 157, 161, 162, 165, 166, 167, 168, 169, 171, 173,174, 176, 177, 178, 179, 180, 182, 183, 184, 186, 187, 188, 189, 192,193, 607, 608, 609, 610, 611 and 614, or a nucleotide sequence derivedfrom the nucleotide sequence by the replacement of u with t according tothe present invention, examples of the shortest variants registered inthe miRBase Release 20 include polynucleotides having sequencesrepresented by SEQ ID NOs: 389, 391, 393, 395, 397, 399, 401, 403, 405,407, 409, 411, 413, 415, 417, 419, 421, 423, 425, 427, 429, 431, 433,435, 437, 439, 441, 443, 445, 447, 449, 451, 453, 455, 457, 459, 461,463, 465, 467, 469, 471, 473, 475, 477, 479, 481, 483, 485, 487, 489,491, 493, 495, 497, 499, 501, 503, 505, 507, 509, 511, 513, 515, 517,519, 521, 523, 525, 527, 529, 531, 533, 535, 537, 539, 541, 543, 545,547, 549, 551, 553, 555, 557, 559, 561, 563, 565, 567, 569, 571, 573,575, 577, 579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601,603, 605, 625, 627, 629, 631, 633 and 635, respectively. In addition tothese variants and fragments, examples thereof include a large number ofisomiR polynucleotides of SEQ ID NOs: 1 to 194 and 606 to 614 registeredin miRBase. Examples of the polynucleotide comprising a nucleotidesequence represented by any of SEQ ID NOs: 1 to 194 and 606 to 614 to494 include a polynucleotide represented by any of SEQ ID NOs: 195 to387 and 615 to 623, which are their respective precursors.

The names and miRBase Accession Nos. (registration numbers) of the genesrepresented by SEQ ID NOs: 1 to 635 are shown in Table 1.

TABLE 1 SEQ miRBase ID NO: Gene name registration No. 1 hsa-miR-6726-5pMIMAT0027353 2 hsa-miR-4257 MIMAT0016878 3 hsa-miR-6787-5p MIMAT00274744 hsa-miR-6780b-5p MIMAT0027572 5 hsa-miR-3131 MIMAT0014996 6hsa-miR-7108-5p MIMAT0028113 7 hsa-miR-1343-3p MIMAT0019776 8hsa-miR-1247-3p MIMAT0022721 9 hsa-miR-4651 MIMAT0019715 10hsa-miR-6757-5p MIMAT0027414 11 hsa-miR-3679-5p MIMAT0018104 12hsa-miR-7641 MIMAT0029782 13 hsa-miR-6746-5p MIMAT0027392 14hsa-miR-8072 MIMAT0030999 15 hsa-miR-6741-5p MIMAT0027383 16hsa-miR-1908-5p MIMAT0007881 17 hsa-miR-6857-5p MIMAT0027614 18hsa-miR-4746-3p MIMAT0019881 19 hsa-miR-744-5p MIMAT0004945 20hsa-miR-4792 MIMAT0019964 21 hsa-miR-564 MIMAT0003228 22 hsa-miR-6791-5pMIMAT0027482 23 hsa-miR-6825-5p MIMAT0027550 24 hsa-miR-6826-5pMIMAT0027552 25 hsa-miR-4665-3p MIMAT0019740 26 hsa-miR-4467MIMAT0018994 27 hsa-miR-3188 MIMAT0015070 28 hsa-miR-6125 MIMAT002459829 hsa-miR-6756-5p MIMAT0027412 30 hsa-miR-1228-3p MIMAT0005583 31hsa-miR-8063 MIMAT0030990 32 hsa-miR-8069 MIMAT0030996 33hsa-miR-6875-5p MIMAT0027650 34 hsa-miR-3185 MIMAT0015065 35 hsa-miR-4433b-3p MIMAT0030414 36 hsa-miR-6887-5p MIMAT0027674 37 hsa-miR-128-1 -5p MIMAT0026477 38 hsa-miR-6724-5p MIMAT0025856 39 hsa-miR-1914-3pMIMAT0007890 40 hsa-miR-1225-5p MIMAT0005572 41 hsa-m iR-4419bMIMAT0019034 42 hsa-miR-7110-5p MIMAT0028117 43 hsa-m iR-187-5pMIMAT0004561 44 hsa-miR-3184-5p MIMAT0015064 45 hsa-m iR-204-3pMIMAT0022693 46 hsa-miR-5572 MIMAT0022260 47 hsa-miR-6729-5pMIMAT0027359 48 hsa-m iR-615-5p MIMAT0004804 49 hsa-miR-6749-5pMIMAT0027398 50 hsa-miR-6515-3p MIMAT0025487 51 hsa-miR-3937MIMAT0018352 52 hsa-miR-6840-3p MIMAT0027583 53 hsa-miR-6893-5pMIMAT0027686 54 hsa-miR-4728-5p MIMAT0019849 55 hsa-miR-6717-5pMIMAT0025846 56 hsa-miR-7113-3p MIMAT0028124 57 hsa-miR-4665-5pMIMAT0019739 58 hsa-miR-642b-3p MIMAT0018444 59 hsa-miR-7109-5pMIMAT0028115 60 hsa-miR-6842-5p MIMAT0027586 61 hsa-miR-4442MIMAT0018960 62 hsa-miR-4433-3p MIMAT0018949 63 hsa-miR-4707-5pMIMAT0019807 64 hsa-miR-6126 MIMAT0024599 65 hsa-miR-4449 MIMAT001896866 hsa-miR-4706 MIMAT0019806 67 hsa-miR-1913 MIMAT0007888 68 hsa-miR-602MIMAT0003270 69 hsa-miR-939-5p MIMAT0004982 70 hsa-miR-4695-5pMIMAT0019788 71 hsa-miR-711 MIMAT0012734 72 hsa-miR-6816-5p MIMAT002753273 hsa-miR-4632-5p MIMAT0022977 74 hsa-miR-6721-5p MIMAT0025852 75hsa-miR-7847-3p MIMAT0030422 76 hsa-miR-6132 MIMAT0024616 77hsa-miR-887-3p MIMAT0004951 78 hsa-miR-3679-3p MIMAT0018105 79hsa-miR-6784-5p MIMAT0027468 80 hsa-miR-1249 MIMAT0005901 81hsa-miR-937-5p MIMAT0022938 82 hsa-miR-5195-3p MIMAT0021127 83hsa-miR-6732-5p MIMAT0027365 84 hsa-miR-4417 MIMAT0018929 85hsa-miR-4281 MIMAT0016907 86 hsa-miR-4734 MIMAT0019859 87hsa-miR-6766-3p MIMAT0027433 88 hsa-miR-663a MIMAT0003326 89hsa-miR-4513 MIMAT0019050 90 hsa-miR-6781 -5p MIMAT0027462 91hsa-miR-1227-5p MIMAT0022941 92 hsa-miR-6845-5p MIMAT0027590 93hsa-miR-6798-5p MIMAT0027496 94 hsa-miR-3620-5p MIMAT0022967 95hsa-miR-1915-5p MIMAT0007891 96 hsa-miR-4294 MIMAT0016849 97hsa-miR-642a-3p MIMAT0020924 98 hsa-miR-371a-5p MIMAT0004687 99hsa-miR-940 MIMAT0004983 100 hsa-miR-4450 MIMAT0018971 101hsa-miR-4723-5p MIMAT0019838 102 hsa-miR-1469 MIMAT0007347 103hsa-miR-6861-5p MIMAT0027623 104 hsa-miR-7975 MIMAT0031178 105hsa-miR-6879-5p MIMAT0027658 106 hsa-miR-6802-5p MIMAT0027504 107hsa-miR- 1268b MIMAT0018925 108 hsa-miR-663b MIMAT0005867 109hsa-miR-125a-3p MIMAT0004602 110 hsa-miR-2861 MIMAT0013802 111hsa-miR-6088 MIMAT0023713 112 hsa-miR-4758-5p MIMAT0019903 113hsa-miR-296-3p MIMAT0004679 114 hsa-miR-6738-5p MIMAT0027377 115hsa-miR-671-5p MIMAT0003880 116 hsa-miR-4454 MIMAT0018976 117hsa-miR-4516 MIMAT0019053 118 hsa-miR-7845-5p MIMAT0030420 119hsa-miR-4741 MIMAT0019871 120 hsa-miR-92b-5p MIMAT0004792 121hsa-miR-6795-5p MIMAT0027490 122 hsa-miR-6805-3p MIMAT0027511 123hsa-miR-4725-3p MIMAT0019844 124 hsa-miR-6782-5p MIMAT0027464 125hsa-miR-4688 MIMAT0019777 126 hsa-miR-6850-5p MIMAT0027600 127hsa-miR-6777-5p MIMAT0027454 128 hsa-miR-6785-5p MIMAT0027470 129hsa-miR-7106-5p MIMAT0028109 130 hsa-miR-3663-3p MIMAT0018085 131hsa-miR-6131 MIMAT0024615 132 hsa-miR-1915-3p MIMAT0007892 133hsa-miR-4532 MIMAT0019071 134 hsa-miR-6820-5p MIMAT0027540 135hsa-miR-4689 MIMAT0019778 136 hsa-miR-4638-5p MIMAT0019695 137hsa-miR-3656 MIMAT0018076 138 hsa-miR-3621 MIMAT0018002 139hsa-miR-6769b-5p MIMAT0027620 140 hsa-miR-149-3p MIMAT0004609 141hsa-miR-23b-3p MIMAT0000418 142 hsa-miR-3135b MIMAT0018985 143hsa-miR-6848-5p MIMAT0027596 144 hsa-miR-6769a-5p MIMAT0027438 145hsa-miR-4327 MIMAT0016889 146 hsa-miR-6765-3p MIMAT0027431 147hsa-miR-6716-5p MIMAT0025844 148 hsa-miR-6877-5p MIMAT0027654 149hsa-miR-6727-5p MIMAT0027355 150 hsa-miR-4534 MIMAT0019073 151hsa-miR-614 MIMAT0003282 152 hsa-miR-1202 MIMAT0005865 153 hsa-miR-575MIMAT0003240 154 hsa-miR-6870-5p MIMAT0027640 155 hsa-miR-6722-3pMIMAT0025854 156 hsa-miR-7977 MIMAT0031180 157 hsa-miR-4649-5pMIMAT0019711 158 hsa-miR-4675 MIMAT0019757 159 hsa-miR-6075 MIMAT0023700160 hsa-miR-6779-5p MIMAT0027458 161 hsa-miR-4271 MIMAT0016901 162hsa-miR-3196 MIMAT0015080 163 hsa-miR-6803-5p MIMAT0027506 164hsa-miR-6789-5p MIMAT0027478 165 hsa-miR-4648 MIMAT0019710 166hsa-miR-4508 MIMAT0019045 167 hsa-miR-4749-5p MIMAT0019885 168hsa-miR-4505 MIMAT0019041 169 hsa-miR-5698 MIMAT0022491 170hsa-miR-1199-5p MIMAT0031119 171 hsa-miR-4763-3p MIMAT0019913 172hsa-miR-1231 MIMAT0005586 173 hsa-miR-1233-5p MIMAT0022943 174hsa-miR-150-3p MIMAT0004610 175 hsa-miR-1225-3p MIMAT0005573 176hsa-miR-92a-2-5p MIMAT0004508 177 hsa-miR-423-5p MIMAT0004748 178hsa-miR-1268a MIMAT0005922 179 hsa-miR-128-2-5p MIMAT0031095 180hsa-miR-24-3p MIMAT0000080 181 hsa-miR-4697-5p MIMAT0019791 182hsa-miR-3197 MIMAT0015082 183 hsa-miR-675-5p MIMAT0004284 184hsa-miR-4486 MIMAT0019020 185 hsa-miR-7107-5p MIMAT0028111 186hsa-miR-23a-3p MIMAT0000078 187 hsa-miR-4667-5p MIMAT0019743 188hsa-miR-451a MIMAT0001631 189 hsa-miR-3940-5p MIMAT0019229 190hsa-miR-8059 MIMAT0030986 191 hsa-miR-6813-5p MIMAT0027526 192hsa-miR-4492 MIMAT0019027 193 hsa-miR-4476 MIMAT0019003 194 hsa-miR-6090MIMAT0023715 195 hsa-mir-6726 MI0022571 196 hsa-mir-4257 MI0015856 197hsa-mir-6787 MI0022632 198 hsa-mir-6780b MI0022681 199 hsa-mir-3131MI0014151 200 hsa-mir-7108 MI0022959 201 hsa-mir-1343 MI0017320 202hsa-mir-1247 MI0006382 203 hsa-mir-4651 MI0017279 204 hsa-mir-6757MI0022602 205 hsa-mir-3679 MI0016080 206 hsa-mir-7641-1 MI0024975 207hsa-mir-7641-2 MI0024976 208 hsa-mir-6746 MI0022591 209 hsa-mir-8072MI0025908 210 hsa-mir-6741 MI0022586 211 hsa-mir-1908 MI0008329 212hsa-mir-6857 MI0022703 213 hsa-mir-4746 MI0017385 214 hsa-mir-744MI0005559 215 hsa-mir-4792 MI0017439 216 hsa-mir-564 MI0003570 217hsa-mir-6791 MI0022636 218 hsa-mir-6825 MI0022670 219 hsa-mir-6826MI0022671 220 hsa-mir-4665 MI0017295 221 hsa-mir-4467 MI0016818 222hsa-mir-3188 MI0014232 223 hsa-mir-6125 MI0021259 224 hsa-mir-6756MI0022601 225 hsa-mir-1228 MI0006318 226 hsa-mir-8063 MI0025899 227hsa-mir-8069 MI0025905 228 hsa-mir-6875 MI0022722 229 hsa-mir-3185MI0014227 230 hsa-mir-4433b MI0025511 231 hsa-mir-6887 MI0022734 232hsa-mir-128-1 MI0000447 233 hsa-mir-6724 MI0022559 234 hsa-mir-1914MI0008335 235 hsa-mir-1225 MI0006311 236 hsa-mir-4419b MI0016861 237hsa-mir-7110 MI0022961 238 hsa-mir-187 MI0000274 239 hsa-mir-3184MI0014226 240 hsa-mir-204 MI0000284 241 hsa-mir-5572 MI0019117 242hsa-mir-6729 MI0022574 243 hsa-mir-615 MI0003628 244 hsa-mir-6749MI0022594 245 hsa-mir-6515 MI0022227 246 hsa-mir-3937 MI0016593 247hsa-mir-6840 MI0022686 248 hsa-mir-6893 MI0022740 249 hsa-mir-4728MI0017365 250 hsa-mir-6717 MI0022551 251 hsa-mir-7113 MI0022964 252hsa-mir-642b MI0016685 253 hsa-mir-7109 MI0022960 254 hsa-mir-6842MI0022688 255 hsa-mir-4442 MI0016785 256 hsa-mir-4433 MI0016773 257hsa-mir-4707 MI0017340 258 hsa-mir-6126 MI0021260 259 hsa-mir-4449MI0016792 260 hsa-mir-4706 MI0017339 261 hsa-mir-1913 MI0008334 262hsa-mir-602 MI0003615 263 hsa-mir-939 MI0005761 264 hsa-mir-4695MI0017328 265 hsa-mir-711 MI0012488 266 hsa-mir-6816 MI0022661 267hsa-mir-4632 MI0017259 268 hsa-mir-6721 MI0022556 269 hsa-mir-7847MI0025517 270 hsa-mir-6132 MI0021277 271 hsa-mir-887 MI0005562 272hsa-mir-6784 MI0022629 273 hsa-mir-1249 MI0006384 274 hsa-mir-937MI0005759 275 hsa-mir-5195 MI0018174 276 hsa-mir-6732 MI0022577 277hsa-mir-4417 MI0016753 278 hsa-mir-4281 MI0015885 279 hsa-mir-4734MI0017371 280 hsa-mir-6766 MI0022611 281 hsa-mir-663a MI0003672 282hsa-mir-4513 MI0016879 283 hsa-mir-6781 MI0022626 284 hsa-mir-1227MI0006316 285 hsa-mir-6845 MI0022691 286 hsa-mir-6798 MI0022643 287hsa-mir-3620 MI0016011 288 hsa-mir-1915 MI0008336 289 hsa-mir-4294MI0015 827 290 hsa-mir-642a MI0003657 291 hsa-mir-371a MI0000779 292hsa-mir-940 MI0005762 293 hsa-mir-4450 MI0016795 294 hsa-mir-4723MI0017359 295 hsa-mir-1469 MI0007074 296 hsa-mir-6861 MI0022708 297hsa-mir-7975 MI0025751 298 hsa-mir-6879 MI0022726 299 hsa-mir-6802MI0022647 300 hsa-mir-1268b MI0016748 301 hsa-mir-663b MI0006336 302hsa-mir-125a MI0000469 303 hsa-mir-2861 MI0013006 304 hsa-mir-6088MI0020365 305 hsa-mir-4758 MI0017399 306 hsa-mir-296 MI0000747 307hsa-mir-6738 MI0022583 308 hsa-mir-671 MI0003760 309 hsa-mir-4454MI0016800 310 hsa-mir-4516 MI0016882 311 hsa-mir-7845 MI0025515 312hsa-mir-4741 MI0017379 313 hsa-mir-92b MI0003560 314 hsa-mir-6795MI0022640 315 hsa-mir-6805 MI0022650 316 hsa-mir-4725 MI0017362 317hsa-mir-6782 MI0022627 318 hsa-mir-4688 MI0017321 319 hsa-mir-6850MI0022696 320 hsa-mir-6777 MI0022622 321 hsa-mir-6785 MI0022630 322hsa-mir-7106 MI0022957 323 hsa-mir-3663 MI0016064 324 hsa-mir-6131MI0021276 325 hsa-mir-4532 MI0016899 326 hsa-mir-6820 MI0022665 327hsa-mir-4689 MI0017322 328 hsa-mir-4638 MI0017265 329 hsa-mir-3656MI0016056 330 hsa-mir-3621 MI0016012 331 hsa-mir-6769b MI0022706 332hsa-mir-149 MI0000478 333 hsa-mir-23b MI0000439 334 hsa-mir-3135bMI0016809 335 hsa-mir-6848 MI0022694 336 hsa-mir- 6769a MI0022614 337hsa-mir-4327 MI0015867 338 hsa-mir-6765 MI0022610 339 hsa-mir-6716MI0022550 340 hsa-mir-6877 MI0022724 341 hsa-mir-6727 MI0022572 342hsa-mir-4534 MI0016901 343 hsa-mir-614 MI0003627 344 hsa-mir-1202MI0006334 345 hsa-mir-575 MI0003582 346 hsa-mir-6870 MI0022717 347hsa-mir-6722 MI0022557 348 hsa-mir-7977 MI0025753 349 hsa-mir-4649MI0017276 350 hsa-mir-4675 MI0017306 351 hsa-mir-6075 MI0020352 352hsa-mir-6779 MI0022624 353 hsa-mir-4271 MI0015879 354 hsa-mir-3196MI0014241 355 hsa-mir-6803 MI0022648 356 hsa-mir-6789 MI0022634 357hsa-mir-4648 MI0017275 358 hsa-mir-4508 MI0016872 359 hsa-mir-4749MI0017388 360 hsa-mir-4505 MI0016868 361 hsa-mir-5698 MI0019305 362hsa-mir-1199 MI0020340 363 hsa-mir-4763 MI0017404 364 hsa-mir-1231MI0006321 365 hsa-mir-1233-1 MI0006323 366 hsa-mir-1233-2 MI0015973 367hsa-mir-150 MI0000479 368 hsa-mir-92a-2 MI0000094 369 hsa-mir-423MI0001445 370 hsa-mir-1268a MI0006405 371 hsa-mir-128-2 MI0000727 372hsa-mir-24-1 MI0000080 373 hsa-mir-24-2 MI0000081 374 hsa-mir-4697MI0017330 375 hsa-mir-3197 MI0014245 376 hsa-mir-675 MI0005416 377hsa-mir-4486 MI0016847 378 hsa-mir-7107 MI0022958 379 hsa-mir-23aMI0000079 380 hsa-mir-4667 MI0017297 381 hsa-mir-45la MI0001729 382hsa-mir-3940 MI0016597 383 hsa-mir-8059 MI0025895 384 hsa-mir-6813MI0022658 385 hsa-mir-4492 MI0016854 386 hsa-mir-4476 MI0016828 387hsa-mir-6090 MI0020367 388 isomiR example 1 of SEQ ID NO: 5 — 389 isomiRexample 2 of SEQ ID NO: 5 — 390 isomiR example 1 of SEQ ID NO: 7 — 391isomiR example 2 of SEQ ID NO: 7 — 392 isomiR example 1 of SEQ ID NO: 8— 393 isomiR example 2 of SEQ ID NO: 8 — 394 isomiR example 1 of SEQ IDNO: 9 — 395 isomiR example 2 of SEQ ID NO: 9 — 396 isomiR example 1 ofSEQ ID NO: 11 — 397 isomiR example 2 of SEQ ID NO: 11 — 398 isomiRexample 1 of SEQ ID NO: 16 — 399 isomiR example 2 of SEQ ID NO: 16 — 400isomiR example 1 of SEQ ID NO: 19 — 401 isomiR example 2 of SEQ ID NO:19 — 402 isomiR example 1 of SEQ ID NO: 20 — 403 isomiR example 2 of SEQID NO: 20 — 404 isomiR example 1 of SEQ ID NO: 21 — 405 isomiR example 2of SEQ ID NO: 21 — 406 isomiR example 1 of SEQ ID NO: 26 — 407 isomiRexample 2 of SEQ ID NO: 26 — 408 isomiR example 1 of SEQ ID NO: 27 — 409isomiR example 2 of SEQ ID NO: 27 — 410 isomiR example 1 of SEQ ID NO:28 — 411 isomiR example 2 of SEQ ID NO: 28 — 412 isomiR example 1 of SEQID NO: 30 — 413 isomiR example 2 of SEQ ID NO: 30 — 414 isomiR example 1of SEQ ID NO: 34 — 415 isomiR example 2 of SEQ ID NO: 34 — 416 isomiRexample 1 of SEQ ID NO: 37 — 417 isomiR example 2 of SEQ ID NO: 37 — 418isomiR example 1 of SEQ ID NO: 38 — 419 isomiR example 2 of SEQ ID NO:38 — 420 isomiR example 1 of SEQ ID NO: 39 — 421 isomiR example 2 of SEQID NO: 39 — 422 isomiR example 1 of SEQ ID NO: 41 — 423 isomiR example 2of SEQ ID NO: 41 — 424 isomiR example 1 of SEQ ID NO: 43 — 425 isomiRexample 2 of SEQ ID NO: 43 — 426 isomiR example 1 of SEQ ID NO: 45 — 427isomiR example 2 of SEQ ID NO: 45 — 428 isomiR example 1 of SEQ ID NO:46 — 429 isomiR example 2 of SEQ ID NO: 46 — 430 isomiR example 1 of SEQID NO: 48 — 431 isomiR example 2 of SEQ ID NO: 48 — 432 isomiR example 1of SEQ ID NO: 50 — 433 isomiR example 2 of SEQ ID NO: 50 — 434 isomiRexample 1 of SEQ ID NO: 54 — 435 isomiR example 2 of SEQ ID NO: 54 — 436isomiR example 1 of SEQ ID NO: 55 — 437 isomiR example 2 of SEQ ID NO:55 — 438 isomiR example 1 of SEQ ID NO: 57 — 439 isomiR example 2 of SEQID NO: 57 — 440 isomiR example 1 of SEQ ID NO: 58 — 441 isomiR example 2of SEQ ID NO: 58 — 442 isomiR example 1 of SEQ ID NO: 61 — 443 isomiRexample 2 of SEQ ID NO: 61 — 444 isomiR example 1 of SEQ ID NO: 62 — 445isomiR example 2 of SEQ ID NO: 62 — 446 isomiR example 1 of SEQ ID NO:63 — 447 isomiR example 2 of SEQ ID NO: 63 — 448 isomiR example 1 of SEQID NO: 64 — 449 isomiR example 2 of SEQ ID NO: 64 — 450 isomiR example 1of SEQ ID NO: 65 — 451 isomiR example 2 of SEQ ID NO: 65 — 452 isomiRexample 1 of SEQ ID NO: 66 — 453 isomiR example 2 of SEQ ID NO: 66 — 454isomiR example 1 of SEQ ID NO: 67 — 455 isomiR example 2 of SEQ ID NO:67 — 456 isomiR example 1 of SEQ ID NO: 69 — 457 isomiR example 2 of SEQID NO: 69 — 458 isomiR example 1 of SEQ ID NO: 70 — 459 isomiR example 2of SEQ ID NO: 70 — 460 isomiR example 1 of SEQ ID NO: 71 — 461 isomiRexample 2 of SEQ ID NO: 71 — 462 isomiR example 1 of SEQ ID NO: 73 — 463isomiR example 2 of SEQ ID NO: 73 — 464 isomiR example 1 of SEQ ID NO:74 — 465 isomiR example 2 of SEQ ID NO: 74 — 466 isomiR example 1 of SEQID NO: 76 — 467 isomiR example 2 of SEQ ID NO: 76 — 468 isomiR example 1of SEQ ID NO: 77 — 469 isomiR example 2 of SEQ ID NO: 77 — 470 isomiRexample 1 of SEQ ID NO: 78 — 471 isomiR example 2 of SEQ ID NO: 78 — 472isomiR example 1 of SEQ ID NO: 80 — 473 isomiR example 2 of SEQ ID NO:80 — 474 isomiR example 1 of SEQ ID NO: 81 — 475 isomiR example 2 of SEQID NO: 81 — 476 isomiR example 1 of SEQ ID NO: 82 — 477 isomiR example 2of SEQ ID NO: 82 — 478 isomiR example 1 of SEQ ID NO: 84 — 479 isomiRexample 2 of SEQ ID NO: 84 — 480 isomiR example 1 of SEQ ID NO: 85 — 481isomiR example 2 of SEQ ID NO: 85 — 482 isomiR example 1 of SEQ ID NO:86 — 483 isomiR example 2 of SEQ ID NO: 86 — 484 isomiR example 1 of SEQID NO: 88 — 485 isomiR example 2 of SEQ ID NO: 88 — 486 isomiR example 1of SEQ ID NO: 89 — 487 isomiR example 2 of SEQ ID NO: 89 — 488 isomiRexample 1 of SEQ ID NO: 94 — 489 isomiR example 2 of SEQ ID NO: 94 — 490isomiR example 1 of SEQ ID NO: 95 — 491 isomiR example 2 of SEQ ID NO:95 — 492 isomiR example 1 of SEQ ID NO: 97 — 493 isomiR example 2 of SEQID NO: 97 — 494 isomiR example 1 of SEQ ID NO: 98 — 495 isomiR example 2of SEQ ID NO: 98 — 496 isomiR example 1 of SEQ ID NO: 99 — 497 isomiRexample 2 of SEQ ID NO: 99 — 498 isomiR example 1 of SEQ ID NO: 100 —499 isomiR example 2 of SEQ ID NO: 100 — 500 isomiR example 1 of SEQ IDNO: 101 — 501 isomiR example 2 of SEQ ID NO: 101 — 502 isomiR example 1of SEQ ID NO: 104 — 503 isomiR example 2 of SEQ ID NO: 104 — 504 isomiRexample 1 of SEQ ID NO: 107 — 505 isomiR example 2 of SEQ ID NO: 107 —506 isomiR example 1 of SEQ ID NO: 108 — 507 isomiR example 2 of SEQ IDNO: 108 — 508 isomiR example 1 of SEQ ID NO: 109 — 509 isomiR example 2of SEQ ID NO: 109 — 510 isomiR example 1 of SEQ ID NO: 110 — 511 isomiRexample 2 of SEQ ID NO: 110 — 512 isomiR example 1 of SEQ ID NO: 111 —513 isomiR example 2 of SEQ ID NO: 111 — 514 isomiR example 1 of SEQ IDNO: 112 — 515 isomiR example 2 of SEQ ID NO: 112 — 516 isomiR example 1of SEQ ID NO: 113 — 517 isomiR example 2 of SEQ ID NO: 113 — 518 isomiRexample 1 of SEQ ID NO: 115 — 519 isomiR example 2 of SEQ ID NO: 115 —520 isomiR example 1 of SEQ ID NO: 116 — 521 isomiR example 2 of SEQ IDNO: 116 — 522 isomiR example 1 of SEQ ID NO: 117 — 523 isomiR example 2of SEQ ID NO: 117 — 524 isomiR example 1 of SEQ ID NO: 119 — 525 isomiRexample 2 of SEQ ID NO: 119 — 526 isomiR example 1 of SEQ ID NO: 120 —527 isomiR example 2 of SEQ ID NO: 120 — 528 isomiR example 1 of SEQ IDNO: 123 — 529 isomiR example 2 of SEQ ID NO: 123 — 530 isomiR example 1of SEQ ID NO: 125 — 531 isomiR example 2 of SEQ ID NO: 125 — 532 isomiRexample 1 of SEQ ID NO: 131 — 533 isomiR example 2 of SEQ ID NO: 131 —534 isomiR example 1 of SEQ ID NO: 132 — 535 isomiR example 2 of SEQ IDNO: 132 — 536 isomiR example 1 of SEQ ID NO: 133 — 537 isomiR example 2of SEQ ID NO: 133 — 538 isomiR example 1 of SEQ ID NO: 135 — 539 isomiRexample 2 of SEQ ID NO: 135 — 540 isomiR example 1 of SEQ ID NO: 136 —541 isomiR example 2 of SEQ ID NO: 136 — 542 isomiR example 1 of SEQ IDNO: 137 — 543 isomiR example 2 of SEQ ID NO: 137 — 544 isomiR example 1of SEQ ID NO: 140 — 545 isomiR example 2 of SEQ ID NO: 140 — 546 isomiRexample 1 of SEQ ID NO: 141 — 547 isomiR example 2 of SEQ ID NO: 141 —548 isomiR example 1 of SEQ ID NO: 142 — 549 isomiR example 2 of SEQ IDNO: 142 — 550 isomiR example 1 of SEQ ID NO: 147 — 551 isomiR example 2of SEQ ID NO: 147 — 552 isomiR example 1 of SEQ ID NO: 151 — 553 isomiRexample 2 of SEQ ID NO: 151 — 554 isomiR example 1 of SEQ ID NO: 152 —555 isomiR example 2 of SEQ ID NO: 152 — 556 isomiR example 1 of SEQ IDNO: 157 — 557 isomiR example 2 of SEQ ID NO: 157 — 558 isomiR example 1of SEQ ID NO: 161 — 559 isomiR example 2 of SEQ ID NO: 161 — 560 isomiRexample 1 of SEQ ID NO: 162 — 561 isomiR example 2 of SEQ ID NO: 162 —562 isomiR example 1 of SEQ ID NO: 165 — 563 isomiR example 2 of SEQ IDNO: 165 — 564 isomiR example 1 of SEQ ID NO: 166 — 565 isomiR example 2of SEQ ID NO: 166 — 566 isomiR example 1 of SEQ ID NO: 167 — 567 isomiRexample 2 of SEQ ID NO: 167 — 568 isomiR example 1 of SEQ ID NO: 168 —569 isomiR example 2 of SEQ ID NO: 168 — 570 isomiR example 1 of SEQ IDNO: 169 — 571 isomiR example 2 of SEQ ID NO: 169 — 572 isomiR example 1of SEQ ID NO: 171 — 573 isomiR example 2 of SEQ ID NO: 171 — 574 isomiRexample 1 of SEQ ID NO: 173 — 575 isomiR example 2 of SEQ ID NO: 173 —576 isomiR example 1 of SEQ ID NO: 174 — 577 isomiR example 2 of SEQ IDNO: 174 — 578 isomiR example 1 of SEQ ID NO: 176 — 579 isomiR example 2of SEQ ID NO: 176 — 580 isomiR example 1 of SEQ ID NO: 177 — 581 isomiRexample 2 of SEQ ID NO: 177 — 582 isomiR example 1 of SEQ ID NO: 178 —583 isomiR example 2 of SEQ ID NO: 178 — 584 isomiR example 1 of SEQ IDNO: 179 — 585 isomiR example 2 of SEQ ID NO: 179 — 586 isomiR example 1of SEQ ID NO: 180 — 587 isomiR example 2 of SEQ ID NO: 180 — 588 isomiRexample 1 of SEQ ID NO: 182 — 589 isomiR example 2 of SEQ ID NO: 182 —590 isomiR example 1 of SEQ ID NO: 183 — 591 isomiR example 2 of SEQ IDNO: 183 — 592 isomiR example 1 of SEQ ID NO: 184 — 593 isomiR example 2of SEQ ID NO: 184 — 594 isomiR example 1 of SEQ ID NO: 186 — 595 isomiRexample 2 of SEQ ID NO: 186 — 596 isomiR example 1 of SEQ ID NO: 187 —597 isomiR example 2 of SEQ ID NO: 187 — 598 isomiR example 1 of SEQ IDNO: 188 — 599 isomiR example 2 of SEQ ID NO: 188 — 600 isomiR example 1of SEQ ID NO: 189 — 601 isomiR example 2 of SEQ ID NO: 189 — 602 isomiRexample 1 of SEQ ID NO: 192 — 603 isomiR example 2 of SEQ ID NO: 192 —604 isomiR example 1 of SEQ ID NO: 193 — 605 isomiR example 2 of SEQ IDNO: 193 — 606 hsa-miR-6836-3p MIMAT0027575 607 hsa-miR-3195 MIMAT0015079608 hsa-miR-718 MIMAT0012735 609 hsa-miR-3178 MIMAT0015055 610hsa-miR-638 MIMAT0003308 611 hsa-miR-4497 MIMAT0019032 612 hsa-miR-6085MIMAT0023710 613 hsa-miR-6752-5p MIMAT0027404 614 hsa-miR-135a-3pMIMAT0004595 615 hsa-mir-6836 MI0022682 616 hsa-mir-3195 MI0014240 617hsa-mir-718 MI0012489 618 hsa-mir-3178 MI0014212 619 hsa-mir-638MI0003653 620 hsa-mir-4497 MI0016859 621 hsa-mir-6085 MI0020362 622hsa-mir-6752 MI0022597 623 hsa-mir-135a MI0000452 624 isomiR example 1of SEQ ID NO: 607 — 625 isomiR example 2 of SEQ ID NO: 607 — 626 isomiRexample 1 of SEQ ID NO: 608 — 627 isomiR example 2 of SEQ ID NO: 608 —628 isomiR example 1 of SEQ ID NO: 609 — 629 isomiR example 2 of SEQ IDNO: 609 — 630 isomiR example 1 of SEQ ID NO: 610 — 631 isomiR example 2of SEQ ID NO: 610 — 632 isomiR example 1 of SEQ ID NO: 611 — 633 isomiRexample 2 of SEQ ID NO: 611 — 634 isomiR example 1 of SEQ ID NO: 614 —635 isomiR example 2 of SEQ ID NO: 614 —

The present specification encompasses the contents described in thespecifications and/or drawings of Japanese Patent Application No.2014-122686 and Japanese Patent Application No. 2015-070182 on which thepriority of the present application is based.

Advantageous Effects of Invention

According to the present invention, colorectal cancer can be detectedeasily and highly accurately.

For example, the presence or absence of colorectal cancer in a patientcan be easily detected by using, as an index, the expression levelmeasurement values of several miRNAs in blood, serum, and/or plasma ofthe patient, which can be collected with limited invasiveness.

BRIEF DESCRIPTION OF DRAWINGS

FIG. 1 This figure shows the relationship between the nucleotidesequences of hsa-miR-3679-5p represented by SEQ ID NO: 11 andhsa-miR-3679-3p represented by SEQ ID NO: 78, which are produced from aprecursor hsa-mir-3679 represented by SEQ ID NO: 205.

FIG. 2 Left diagram: the expression level measurement values ofhsa-miR-6726-5p (SEQ ID NO: 1) in healthy subjects (100 persons) andcolorectal cancer patients (34 persons) selected as a training cohortwere each plotted on the ordinate. The horizontal line in the diagramdepicts a threshold (9.43) that was optimized by Fisher's discriminantanalysis and discriminated between the two groups. Right diagram: theexpression level measurement values of hsa-miR-6726-5p (SEQ ID NO: 1) inhealthy subjects (50 persons) and colorectal cancer patients (16persons) selected as validation cohort were each plotted on theordinate. The horizontal line in the diagram depicts the threshold(9.43) that was set for the training cohort and discriminated betweenthe two groups.

FIG. 3 Left diagram: the expression level measurement values ofhsa-miR-6726-5p (SEQ ID NO: 1) in healthy subjects (100 persons,circles) and colorectal cancer patients (34 persons, triangles) selectedas a training cohort were each plotted on the abscissa against theirexpression level measurement values of hsa-miR-4257 (SEQ ID NO: 2) onthe ordinate. The line in the diagram depicts a discriminant function(0=1.26x+y−18.06) that was optimized by Fisher's discriminant analysisand discriminated between the two groups. Right diagram: the expressionlevel measurement values of hsa-miR-6726-5p (SEQ ID NO: 1) in healthysubjects (50 persons, circles) and colorectal cancer patients (16persons, triangles) selected as a validation cohort were each plotted onthe abscissa against their expression level measurement values ofhsa-miR-4257 (SEQ ID NO: 2) on the ordinate. The line in the diagramdepicts the threshold (0=1.26x+y−18.06) that was set in the trainingcohort and discriminated between the two groups.

FIG. 4 Upper diagram: a discriminant(1.49×hsa-miR-3131−0.23×hsa-miR-7847-3p−1.13×hsa-miR-3196+1.11×hsa-miR-3195+2.25×hsa-miR-4665-5p−1.00×hsa-miR-204−3p−11.16)was prepared by use of Fisher's discriminant analysis from theexpression level measurement values of hsa-miR-3131 (SEQ ID NO: 5),hsa-miR-204-3p (SEQ ID NO: 45), hsa-miR-4665-5p (SEQ ID NO: 57),hsa-miR-7847-3p (SEQ ID NO: 75), hsa-miR-3196 (SEQ ID NO: 162), andhsa-miR-3195 (SEQ ID NO: 607) in 34 colorectal cancer patients, 103healthy subjects, 69 pancreatic cancer patients, 66 bile duct cancerpatients, 30 stomach cancer patients, 33 esophageal cancer patients, 32liver cancer patients, and 15 benign pancreaticobiliary disease patientsselected as a training cohort, and discriminant scores obtained from thediscriminant were plotted on the ordinate against the sample groups onthe abscissa. The dotted line in the diagram depicts a discriminantboundary that offered a discriminant score of 0 and discriminatedbetween the groups. Lower diagram: discriminant scores obtained from thediscriminant prepared in the training cohort as to the expression levelmeasurement values of hsa-miR-3131 (SEQ ID NO: 5), hsa-miR-204-3p (SEQID NO: 45), hsa-miR-4665-5p (SEQ ID NO: 57), hsa-miR-7847-3p (SEQ ID NO:75), hsa-miR-3196 (SEQ ID NO: 162), and hsa-miR-3195 (SEQ ID NO: 607) in16 colorectal cancer patients, 47 healthy subjects, 30 pancreatic cancerpatients, 33 bile duct cancer patients, 20 stomach cancer patients, 17esophageal cancer patients, 20 liver cancer patients, and 6 benignpancreaticobiliary disease patients selected as a validation cohort wereplotted on the ordinate against the sample groups on the abscissa. Thedotted line in the diagram depicts the discriminant boundary thatoffered a discriminant score of 0 and discriminated between the twogroups.

DESCRIPTION OF EMBODIMENTS

Hereinafter, the present invention will be described furtherspecifically.

1. Target Nucleic Acid for Colorectal Cancer

A primary target nucleic acid as a colorectal cancer marker fordetecting the presence and/or absence of colorectal cancer or colorectalcancer cells using the nucleic acid probe or the primer for thedetection of colorectal cancer defined above according to the presentinvention can use at least one or more miRNA(s) selected from the groupconsisting of hsa-miR-6726-5p, hsa-miR-4257, hsa-miR-6787-5p,hsa-miR-6780b-5p, hsa-miR-3131, hsa-miR-7108-5p, hsa-miR-1343-3p,hsa-miR-1247-3p, hsa-miR-4651, hsa-miR-6757-5p, hsa-miR-3679-5p,hsa-miR-7641, hsa-miR-6746-5p, hsa-miR-8072, hsa-miR-6741-5p,hsa-miR-1908-5p, hsa-miR-6857-5p, hsa-miR-4746-3p, hsa-miR-744-5p,hsa-miR-4792, hsa-miR-564, hsa-miR-6791-5p, hsa-miR-6825-5p,hsa-miR-6826-5p, hsa-miR-4665-3p, hsa-miR-4467, hsa-miR-3188,hsa-miR-6125, hsa-miR-6756-5p, hsa-miR-1228-3p, hsa-miR-8063,hsa-miR-8069, hsa-miR-6875-5p, hsa-miR-3185, hsa-miR-4433b-3p,hsa-miR-6887-5p, hsa-miR-128-1-5p, hsa-miR-6724-5p, hsa-miR-1914-3p,hsa-miR-1225-5p, hsa-miR-4419b, hsa-miR-7110-5p, hsa-miR-187-5p,hsa-miR-3184-5p, hsa-miR-204-3p, hsa-miR-5572, hsa-miR-6729-5p,hsa-miR-615-5p, hsa-miR-6749-5p, hsa-miR-6515-3p, hsa-miR-3937,hsa-miR-6840-3p, hsa-miR-6893-5p, hsa-miR-4728-5p, hsa-miR-6717-5p,hsa-miR-7113-3p, hsa-miR-4665-5p, hsa-miR-642b-3p, hsa-miR-7109-5p,hsa-miR-6842-5p, hsa-miR-4442, hsa-miR-4433-3p, hsa-miR-4707-5p,hsa-miR-6126, hsa-miR-4449, hsa-miR-4706, hsa-miR-1913, hsa-miR-602,hsa-miR-939-5p, hsa-miR-4695-5p, hsa-miR-711, hsa-miR-6816-5p,hsa-miR-4632-5p, hsa-miR-6721-5p, hsa-miR-7847-3p, hsa-miR-6132,hsa-miR-887-3p, hsa-miR-3679-3p, hsa-miR-6784-5p, hsa-miR-1249,hsa-miR-937-5p, hsa-miR-5195-3p, hsa-miR-6732-5p, hsa-miR-4417,hsa-miR-4281, hsa-miR-4734, hsa-miR-6766-3p, hsa-miR-663a, hsa-miR-4513,hsa-miR-6781-5p, hsa-miR-1227-5p, hsa-miR-6845-5p, hsa-miR-6798-5p,hsa-miR-3620-5p, hsa-miR-1915-5p, hsa-miR-4294, hsa-miR-642a-3p,hsa-miR-371a-5p, hsa-miR-940, hsa-miR-4450, hsa-miR-4723-5p,hsa-miR-1469, hsa-miR-6861-5p, hsa-miR-7975, hsa-miR-6879-5p,hsa-miR-6802-5p, hsa-miR-1268b, hsa-miR-663b, hsa-miR-125a-3p,hsa-miR-2861, hsa-miR-6088, hsa-miR-4758-5p, hsa-miR-296-3p,hsa-miR-6738-5p, hsa-miR-671-5p, hsa-miR-4454, hsa-miR-4516,hsa-miR-7845-5p, hsa-miR-4741, hsa-miR-92b-5p, hsa-miR-6795-5p,hsa-miR-6805-3p, hsa-miR-4725-3p, hsa-miR-6782-5p, hsa-miR-4688,hsa-miR-6850-5p, hsa-miR-6777-5p, hsa-miR-6785-5p, hsa-miR-7106-5p,hsa-miR-3663-3p, hsa-miR-6131, hsa-miR-1915-3p, hsa-miR-4532,hsa-miR-6820-5p, hsa-miR-4689, hsa-miR-4638-5p, hsa-miR-3656,hsa-miR-3621, hsa-miR-6769b-5p, hsa-miR-149-3p, hsa-miR-23b-3p,hsa-miR-3135b, hsa-miR-6848-5p, hsa-miR-6769a-5p, hsa-miR-4327,hsa-miR-6765-3p, hsa-miR-6716-5p, hsa-miR-68T7-5p, hsa-miR-6727-5p,hsa-miR-4534, hsa-miR-614, hsa-miR-1202, hsa-miR-575, hsa-miR-6870-5p,hsa-miR-6722-3p, hsa-miR-7977, hsa-miR-4649-5p, hsa-miR-4675,hsa-miR-6075, hsa-miR-6779-5p, hsa-miR-4271, hsa-miR-3196,hsa-miR-6803-5p, hsa-miR-6789-5p, hsa-miR-4648, hsa-miR-4508,hsa-miR-4749-5p, hsa-miR-4505, hsa-miR-5698, hsa-miR-1199-5p,hsa-miR-4763-3p, hsa-miR-6836-3p, hsa-miR-3195, hsa-miR-718,hsa-miR-3178, hsa-miR-638, hsa-miR-4497, hsa-miR-6085, hsa-miR-6752-5pand hsa-miR-135a-3p. Furthermore, at least one or more miRNA(s) selectedfrom the group consisting of other colorectal cancer markers that can becombined with these miRNAs, i.e., hsa-miR-1231, hsa-miR-1233-5p,hsa-miR-150-3p, hsa-miR-1225-3p, hsa-miR-92a-2-5p, hsa-miR-423-5p,hsa-miR-1268a, hsa-miR-128-2-5p and hsa-miR-24-3p can also be preferablyused as a target nucleic acid. Moreover, at least one or more miRNA(s)selected from the group consisting of other colorectal cancer markersthat can be combined with these miRNAs, i.e., hsa-miR-4697-5p,hsa-miR-3197, hsa-miR-675-5p, hsa-miR-4486, hsa-miR-7107-5p,hsa-miR-23a-3p, hsa-miR-4667-5p, hsa-miR-451a, hsa-miR-3940-5p,hsa-miR-8059, hsa-miR-6813-5p, hsa-miR-4492, hsa-miR-4476 andhsa-miR-6090 can also be preferably used as a target nucleic acid.

These miRNAs include, for example, a human gene comprising a nucleotidesequence represented by any of SEQ ID NOs: 1 to 194 and 606 to 614(i.e., hsa-miR-6726-5p, hsa-miR-4257, hsa-miR-6787-5p, hsa-miR-6780b-5p,hsa-miR-3131, hsa-miR-7108-5p, hsa-miR-1343-3p, hsa-miR-1247-3p,hsa-miR-4651, hsa-miR-6757-5p, hsa-miR-3679-5p, hsa-miR-7641,hsa-miR-6746-5p, hsa-miR-8072, hsa-miR-6741-5p, hsa-miR-1908-5p,hsa-miR-6857-5p, hsa-miR-4746-3p, hsa-miR-744-5p, hsa-miR-4792,hsa-miR-564, hsa-miR-6791-5p, hsa-miR-6825-5p, hsa-miR-6826-5p,hsa-miR-4665-3p, hsa-miR-4467, hsa-miR-3188, hsa-miR-6125,hsa-miR-6756-5p, hsa-miR-1228-3p, hsa-miR-8063, hsa-miR-8069,hsa-miR-6875-5p, hsa-miR-3185, hsa-miR-4433b-3p, hsa-miR-6887-5p,hsa-miR-128-1-5p, hsa-miR-6724-5p, hsa-miR-1914-3p, hsa-miR-1225-5p,hsa-miR-4419b, hsa-miR-7110-5p, hsa-miR-187-5p, hsa-miR-3184-5p,hsa-miR-204-3p, hsa-miR-5572, hsa-miR-6729-5p, hsa-miR-615-5p,hsa-miR-6749-5p, hsa-miR-6515-3p, hsa-miR-3937, hsa-miR-6840-3p,hsa-miR-6893-5p, hsa-miR-4728-5p, hsa-miR-6717-5p, hsa-miR-7113-3p,hsa-miR-4665-5p, hsa-miR-642b-3p, hsa-miR-7109-5p, hsa-miR-6842-5p,hsa-miR-4442, hsa-miR-4433-3p, hsa-miR-4707-5p, hsa-miR-6126,hsa-miR-4449, hsa-miR-4706, hsa-miR-1913, hsa-miR-602, hsa-miR-939-5p,hsa-miR-4695-5p, hsa-miR-711, hsa-miR-6816-5p, hsa-miR-4632-5p,hsa-miR-6721-5p, hsa-miR-7847-3p, hsa-miR-6132, hsa-miR-887-3p,hsa-miR-3679-3p, hsa-miR-6784-5p, hsa-miR-1249, hsa-miR-937-5p,hsa-miR-5195-3p, hsa-miR-6732-5p, hsa-miR-4417, hsa-miR-4281,hsa-miR-4734, hsa-miR-6766-3p, hsa-miR-663a, hsa-miR-4513,hsa-miR-6781-5p, hsa-miR-1227-5p, hsa-miR-6845-5p, hsa-miR-6798-5p,hsa-miR-3620-5p, hsa-miR-1915-5p, hsa-miR-4294, hsa-miR-642a-3p,hsa-miR-371a-5p, hsa-miR-940, hsa-miR-4450, hsa-miR-4723-5p,hsa-miR-1469, hsa-miR-6861-5p, hsa-miR-7975, hsa-miR-6879-5p,hsa-miR-6802-5p, hsa-miR-1268b, hsa-miR-663b, hsa-miR-125a-3p,hsa-miR-2861, hsa-miR-6088, hsa-miR-4758-5p, hsa-miR-296-3p,hsa-miR-6738-5p, hsa-miR-671-5p, hsa-miR-4454, hsa-miR-4516,hsa-miR-7845-5p, hsa-miR-4741, hsa-miR-92b-5p, hsa-miR-6795-5p,hsa-miR-6805-3p, hsa-miR-4725-3p, hsa-miR-6782-5p, hsa-miR-4688,hsa-miR-6850-5p, hsa-miR-6777-5p, hsa-miR-6785-5p, hsa-miR-7106-5p,hsa-miR-3663-3p, hsa-miR-6131, hsa-miR-1915-3p, hsa-miR-4532,hsa-miR-6820-5p, hsa-miR-4689, hsa-miR-4638-5p, hsa-miR-3656,hsa-miR-3621, hsa-miR-6769b-5p, hsa-miR-149-3p, hsa-miR-23b-3p,hsa-miR-3135b, hsa-miR-6848-5p, hsa-miR-6769a-5p, hsa-miR-4327,hsa-miR-6765-3p, hsa-miR-6716-5p, hsa-miR-6877-5p, hsa-miR-6727-5p,hsa-miR-4534, hsa-miR-614, hsa-miR-1202, hsa-miR-575, hsa-miR-6870-5p,hsa-miR-6722-3p, hsa-miR-7977, hsa-miR-4649-5p, hsa-miR-4675,hsa-miR-6075, hsa-miR-6779-5p, hsa-miR-4271, hsa-miR-3196,hsa-miR-6803-5p, hsa-miR-6789-5p, hsa-miR-4648, hsa-miR-4508,hsa-miR-4749-5p, hsa-miR-4505, hsa-miR-5698, hsa-miR-1199-5p,hsa-miR-4763-3p, hsa-miR-1231, hsa-miR-1233-5p, hsa-miR-150-3p,hsa-miR-1225-3p, hsa-miR-92a-2-5p, hsa-miR-423-5p, hsa-miR-1268a,hsa-miR-128-2-5p, hsa-miR-24-3p, hsa-miR-4697-5p, hsa-miR-3197,hsa-miR-675-5p, hsa-miR-4486, hsa-miR-7107-5p, hsa-miR-23a-3p,hsa-miR-4667-5p, hsa-miR-451a, hsa-miR-3940-5p, hsa-miR-8059,hsa-miR-6813-5p, hsa-miR-4492, hsa-miR-4476, hsa-miR-6090,hsa-miR-6836-3p, hsa-miR-3195, hsa-miR-718, hsa-miR-3178, hsa-miR-638,hsa-miR-4497, hsa-miR-6085, hsa-miR-6752-5p and hsa-miR-135a-3p,respectively), a congener thereof, a transcript thereof, and a variantor a derivative thereof. In this context, the gene, the congener, thetranscript, the variant, and the derivative are as defined above.

The target nucleic acid is preferably a human gene comprising anucleotide sequence represented by any of SEQ ID NOs: 1 to 635 or atranscript thereof, more preferably the transcript, i.e., a miRNA or itsprecursor RNA (pri-miRNA or pre-miRNA).

The first target gene is the hsa-miR-6726-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The second target gene is the hsa-miR-4257 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The third target gene is the hsa-miR-6787-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The fourth target gene is the hsa-miR-6780b-5p gene, a congener thereof,a transcript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The fifth target gene is the hsa-miR-3131 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The sixth target gene is the hsa-miR-7108-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The seventh target gene is the hsa-miR-1343-3p gene, a congener thereof,a transcript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The eighth target gene is the hsa-miR-1247-3p gene, a congener thereof,a transcript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The ninth target gene is the hsa-miR-4651 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 10th target gene is the hsa-miR-6757-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 11th target gene is the hsa-miR-3679-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 12th target gene is the hsa-miR-7641 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 13th target gene is the hsa-miR-6746-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 14th target gene is the hsa-miR-8072 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 15th target gene is the hsa-miR-6741-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 16th target gene is the hsa-miR-1908-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 17th target gene is the hsa-miR-6857-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 18th target gene is the hsa-miR-4746-3p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 19th target gene is the hsa-miR-744-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 20th target gene is the hsa-miR-4792 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 21st target gene is the hsa-miR-564 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 22nd target gene is the hsa-miR-6791-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 23rd target gene is the hsa-miR-6825-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 24th target gene is the hsa-miR-6826-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 25th target gene is the hsa-miR-4665-3p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 26th target gene is the hsa-miR-4467 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 27th target gene is the hsa-miR-3188 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 28th target gene is the hsa-miR-6125 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 29th target gene is the hsa-miR-6756-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 30th target gene is the hsa-miR-1228-3p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 31st target gene is the hsa-miR-8063 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 32nd target gene is the hsa-miR-8069 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 33rd target gene is the hsa-miR-6875-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 34th target gene is the hsa-miR-3185 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 35th target gene is the hsa-miR-4433b-3p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 36th target gene is the hsa-miR-6887-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 37th target gene is the hsa-miR-128-1-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 38th target gene is the hsa-miR-6724-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 39th target gene is the hsa-miR-1914-3p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 40th target gene is the hsa-miR-1225-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 41st target gene is the hsa-miR-4419b gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 42nd target gene is the hsa-miR-7110-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 43rd target gene is the hsa-miR-187-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 44th target gene is the hsa-miR-3184-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 45th target gene is the hsa-miR-204-3p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 46th target gene is the hsa-miR-5572 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 47th target gene is the hsa-miR-6729-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 48th target gene is the hsa-miR-615-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 49th target gene is the hsa-miR-6749-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 50th target gene is the hsa-miR-6515-3p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 51st target gene is the hsa-miR-3937 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 52nd target gene is the hsa-miR-6840-3p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 53rd target gene is the hsa-miR-6893-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 54th target gene is the hsa-miR-4728-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 55th target gene is the hsa-miR-6717-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 56th target gene is the hsa-miR-7113-3p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 57th target gene is the hsa-miR-4665-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 58th target gene is the hsa-miR-642b-3p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 59th target gene is the hsa-miR-7109-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 60th target gene is the hsa-miR-6842-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 61st target gene is the hsa-miR-4442 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 62nd target gene is the hsa-miR-4433-3p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 63rd target gene is the hsa-miR-4707-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 64th target gene is the hsa-miR-6126 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 65th target gene is the hsa-miR-4449 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 66th target gene is the hsa-miR-4706 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 67th target gene is the hsa-miR-1913 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 68th target gene is the hsa-miR-602 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 69th target gene is the hsa-miR-939-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 70th target gene is the hsa-miR-4695-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 71st target gene is the hsa-miR-711 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 72nd target gene is the hsa-miR-6816-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 73rd target gene is the hsa-miR-4632-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 74th target gene is the hsa-miR-6721-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 75th target gene is the hsa-miR-7847-3p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 76th target gene is the hsa-miR-6132 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 77th target gene is the hsa-miR-887-3p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 78th target gene is the hsa-miR-3679-3p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 79th target gene is the hsa-miR-6784-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 80th target gene is the hsa-miR-1249 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 81st target gene is the hsa-miR-937-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 82nd target gene is the hsa-miR-5195-3p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 83rd target gene is the hsa-miR-6732-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 84th target gene is the hsa-miR-4417 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 85th target gene is the hsa-miR-4281 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 86th target gene is the hsa-miR-4734 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 87th target gene is the hsa-miR-6766-3p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 88th target gene is the hsa-miR-663a gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 89th target gene is the hsa-miR-4513 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 90th target gene is the hsa-miR-6781-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 91st target gene is the hsa-miR-1227-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 92nd target gene is the hsa-miR-6845-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 93rd target gene is the hsa-miR-6798-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 94th target gene is the hsa-miR-3620-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 95th target gene is the hsa-miR-1915-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 96th target gene is the hsa-miR-4294 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 97th target gene is the hsa-miR-642a-3p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 98th target gene is the hsa-miR-371a-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 99th target gene is the hsa-miR-940 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 100th target gene is the hsa-miR-4450 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 101st target gene is the hsa-miR-4723-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 102nd target gene is the hsa-miR-1469 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 103rd target gene is the hsa-miR-6861-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 104th target gene is the hsa-miR-7975 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 105th target gene is the hsa-miR-6879-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 106th target gene is the hsa-miR-6802-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 107th target gene is the hsa-miR-1268b gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 108th target gene is the hsa-miR-663b gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 109th target gene is the hsa-miR-125a-3p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 110th target gene is the hsa-miR-2861 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 111th target gene is the hsa-miR-6088 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 112th target gene is the hsa-miR-4758-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 113th target gene is the hsa-miR-296-3p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 114th target gene is the hsa-miR-6738-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 115th target gene is the hsa-miR-671-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 116th target gene is the hsa-miR-4454 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 117th target gene is the hsa-miR-4516 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 118th target gene is the hsa-miR-7845-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 119th target gene is the hsa-miR-4741 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 120th target gene is the hsa-miR-92b-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 121st target gene is the hsa-miR-6795-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 122nd target gene is the hsa-miR-6805-3p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 123rd target gene is the hsa-miR-4725-3p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 124th target gene is the hsa-miR-6782-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 125th target gene is the hsa-miR-4688 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 126th target gene is the hsa-miR-6850-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 127th target gene is the hsa-miR-6777-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 128th target gene is the hsa-miR-6785-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 129th target gene is the hsa-miR-7106-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 130th target gene is the hsa-miR-3663-3p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 131st target gene is the hsa-miR-6131 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 132nd target gene is the hsa-miR-1915-3p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 133rd target gene is the hsa-miR-4532 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 134th target gene is the hsa-miR-6820-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 135th target gene is the hsa-miR-4689 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 136th target gene is the hsa-miR-4638-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 137th target gene is the hsa-miR-3656 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 138th target gene is the hsa-miR-3621 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 139th target gene is the hsa-miR-6769b-5p gene, a congener thereof,a transcript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 140th target gene is the hsa-miR-149-3p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 141st target gene is the hsa-miR-23b-3p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 142nd target gene is the hsa-miR-3135b gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 143rd target gene is the hsa-miR-6848-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 144th target gene is the hsa-miR-6769a-5p gene, a congener thereof,a transcript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 145th target gene is the hsa-miR-4327 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 146th target gene is the hsa-miR-6765-3p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 147th target gene is the hsa-miR-6716-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 148th target gene is the hsa-miR-6877-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 149th target gene is the hsa-miR-6727-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 150th target gene is the hsa-miR-4534 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 151st target gene is the hsa-miR-614 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 152nd target gene is the hsa-miR-1202 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 153rd target gene is the hsa-miR-575 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 154th target gene is the hsa-miR-6870-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 155th target gene is the hsa-miR-6722-3p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 156th target gene is the hsa-miR-7977 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 157th target gene is the hsa-miR-4649-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 158th target gene is the hsa-miR-4675 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 159th target gene is the hsa-miR-6075 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 160th target gene is the hsa-miR-6779-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 161st target gene is the hsa-miR-4271 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 162nd target gene is the hsa-miR-3196 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 163rd target gene is the hsa-miR-6803-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 164th target gene is the hsa-miR-6789-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 165th target gene is the hsa-miR-4648 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 166th target gene is the hsa-miR-4508 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 167th target gene is the hsa-miR-4749-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 168th target gene is the hsa-miR-4505 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 169th target gene is the hsa-miR-5698 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 170th target gene is the hsa-miR-1199-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 171st target gene is the hsa-miR-4763-3p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 172nd target gene is the hsa-miR-1231 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. The previouslyknown report shows that change in the expression of the gene or thetranscript thereof can serve as a marker for colorectal cancer (PatentLiterature 3).

The 173rd target gene is the hsa-miR-1233-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. The previouslyknown report shows that change in the expression of the gene or thetranscript thereof can serve as a marker for colorectal cancer (PatentLiterature 2).

The 174th target gene is the hsa-miR-150-3p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. The previouslyknown report shows that change in the expression of the gene or thetranscript thereof can serve as a marker for colorectal cancer (PatentLiterature 4).

The 175th target gene is the hsa-miR-1225-3p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. The previouslyknown report shows that change in the expression of the gene or thetranscript thereof can serve as a marker for colorectal cancer (PatentLiterature 2).

The 176th target gene is the hsa-miR-92a-2-5p gene, a congener thereof,a transcript thereof, or a variant or a derivative thereof. Thepreviously known report shows that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer(Patent Literatures 1 and 4).

The 177th target gene is the hsa-miR-423-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. The previouslyknown report shows that change in the expression of the gene or thetranscript thereof can serve as a marker for colorectal cancer (PatentLiterature 3).

The 178th target gene is the hsa-miR-1268a gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. The previouslyknown report shows that change in the expression of the gene or thetranscript thereof can serve as a marker for colorectal cancer (PatentLiterature 3).

The 179th target gene is the hsa-miR-128-2-5p gene, a congener thereof,a transcript thereof, or a variant or a derivative thereof. Thepreviously known report shows that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer(Patent Literature 1).

The 180th target gene is the hsa-miR-24-3p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. The previouslyknown report shows that change in the expression of the gene or thetranscript thereof can serve as a marker for colorectal cancer (PatentLiterature 1).

The 181st target gene is the hsa-miR-4697-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 182nd target gene is the hsa-miR-3197 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 183rd target gene is the hsa-miR-675-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 184th target gene is the hsa-miR-4486 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 185th target gene is the hsa-miR-7107-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 186th target gene is the hsa-miR-23a-3p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. The previouslyknown report shows that change in the expression of the gene or thetranscript thereof can serve as a marker for colorectal cancer (PatentLiterature 2).

The 187th target gene is the hsa-miR-4667-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 188th target gene is the hsa-miR-451a gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 189th target gene is the hsa-miR-3940-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 190th target gene is the hsa-miR-8059 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 191st target gene is the hsa-miR-6813-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 192nd target gene is the hsa-miR-4492 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 193rd target gene is the hsa-miR-4476 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 194th target gene is the hsa-miR-6090 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 195th target gene is the hsa-miR-6836-3p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 196th target gene is the hsa-miR-3195 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 197th target gene is the hsa-miR-718 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 198th target gene is the hsa-miR-3178 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 199th target gene is the hsa-miR-638 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 200th target gene is the hsa-miR-4497 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 201st target gene is the hsa-miR-6085 gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 202nd target gene is the hsa-miR-6752-5p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

The 203rd target gene is the hsa-miR-135a-3p gene, a congener thereof, atranscript thereof, or a variant or a derivative thereof. None of thepreviously known reports show that change in the expression of the geneor the transcript thereof can serve as a marker for colorectal cancer.

2. Nucleic Acid Probe or Primer for Detection of Colorectal Cancer

In the present invention, a nucleic acid capable of specifically bindingto any of the target nucleic acids as the colorectal cancer markersdescribed above can be used as a nucleic acid, for example, a nucleicacid probe or a primer, for the detection or diagnosis of colorectalcancer.

In the present invention, the nucleic acid probe or the primer that canbe used for detecting colorectal cancer or for diagnosing colorectalcancer permits qualitative and/or quantitative measurement of thepresence, expression level, or abundance of any of the target nucleicacids as the colorectal cancer markers described above, for example,human-derived hsa-miR-6726-5p, hsa-miR-4257, hsa-miR-6787-5p,hsa-miR-6780b-5p, hsa-miR-3131, hsa-miR-7108-5p, hsa-miR-1343-3p,hsa-miR-1247-3p, hsa-miR-4651, hsa-miR-6757-5p, hsa-miR-3679-5p,hsa-miR-7641, hsa-miR-6746-5p, hsa-miR-8072, hsa-miR-6741-5p,hsa-miR-1908-5p, hsa-miR-6857-5p, hsa-miR-4746-3p, hsa-miR-744-5p,hsa-miR-4792, hsa-miR-564, hsa-miR-6791-5p, hsa-miR-6825-5p,hsa-miR-6826-5p, hsa-miR-4665-3p, hsa-miR-4467, hsa-miR-3188,hsa-miR-6125, hsa-miR-6756-5p, hsa-miR-1228-3p, hsa-miR-8063,hsa-miR-8069, hsa-miR-6875-5p, hsa-miR-3185, hsa-miR-4433b-3p,hsa-miR-6887-5p, hsa-miR-128-1-5p, hsa-miR-6724-5p, hsa-miR-1914-3p,hsa-miR-1225-5p, hsa-miR-4419b, hsa-miR-7110-5p, hsa-miR-187-5p,hsa-miR-3184-5p, hsa-miR-204-3p, hsa-miR-5572, hsa-miR-6729-5p,hsa-miR-615-5p, hsa-miR-6749-5p, hsa-miR-6515-3p, hsa-miR-3937,hsa-miR-6840-3p, hsa-miR-6893-5p, hsa-miR-4728-5p, hsa-miR-6717-5p,hsa-miR-7113-3p, hsa-miR-4665-5p, hsa-miR-642b-3p, hsa-miR-7109-5p,hsa-miR-6842-5p, hsa-miR-4442, hsa-miR-4433-3p, hsa-miR-4707-5p,hsa-miR-6126, hsa-miR-4449, hsa-miR-4706, hsa-miR-1913, hsa-miR-602,hsa-miR-939-5p, hsa-miR-4695-5p, hsa-miR-711, hsa-miR-6816-5p,hsa-miR-4632-5p, hsa-miR-6721-5p, hsa-miR-7847-3p, hsa-miR-6132,hsa-miR-887-3p, hsa-miR-3679-3p, hsa-miR-6784-5p, hsa-miR-1249,hsa-miR-937-5p, hsa-miR-5195-3p, hsa-miR-6732-5p, hsa-miR-4417,hsa-miR-4281, hsa-miR-4734, hsa-miR-6766-3p, hsa-miR-663a, hsa-miR-4513,hsa-miR-6781-5p, hsa-miR-1227-5p, hsa-miR-6845-5p, hsa-miR-6798-5p,hsa-miR-3620-5p, hsa-miR-1915-5p, hsa-miR-4294, hsa-miR-642a-3p,hsa-miR-371a-5p, hsa-miR-940, hsa-miR-4450, hsa-miR-4723-5p,hsa-miR-1469, hsa-miR-6861-5p, hsa-miR-7975, hsa-miR-6879-5p,hsa-miR-6802-5p, hsa-miR-1268b, hsa-miR-663b, hsa-miR-125a-3p,hsa-miR-2861, hsa-miR-6088, hsa-miR-4758-5p, hsa-miR-296-3p,hsa-miR-6738-5p, hsa-miR-671-5p, hsa-miR-4454, hsa-miR-4516,hsa-miR-7845-5p, hsa-miR-4741, hsa-miR-92b-5p, hsa-miR-6795-5p,hsa-miR-6805-3p, hsa-miR-4725-3p, hsa-miR-6782-5p, hsa-miR-4688,hsa-miR-6850-5p, hsa-miR-6777-5p, hsa-miR-6785-5p, hsa-miR-7106-5p,hsa-miR-3663-3p, hsa-miR-6131, hsa-miR-1915-3p, hsa-miR-4532,hsa-miR-6820-5p, hsa-miR-4689, hsa-miR-4638-5p, hsa-miR-3656,hsa-miR-3621, hsa-miR-6769b-5p, hsa-miR-149-3p, hsa-miR-23b-3p,hsa-miR-3135b, hsa-miR-6848-5p, hsa-miR-6769a-5p, hsa-miR-4327,hsa-miR-6765-3p, hsa-miR-6716-5p, hsa-miR-6877-5p, hsa-miR-6727-5p,hsa-miR-4534, hsa-miR-614, hsa-miR-1202, hsa-miR-575, hsa-miR-6870-5p,hsa-miR-6722-3p, hsa-miR-7977, hsa-miR-4649-5p, hsa-miR-4675,hsa-miR-6075, hsa-miR-6779-5p, hsa-miR-4271, hsa-miR-3196,hsa-miR-6803-5p, hsa-miR-6789-5p, hsa-miR-4648, hsa-miR-4508,hsa-miR-4749-5p, hsa-miR-4505, hsa-miR-5698, hsa-miR-1199-5p,hsa-miR-4763-3p, hsa-miR-6836-3p, hsa-miR-3195, hsa-miR-718,hsa-miR-3178, hsa-miR-638, hsa-miR-4497, hsa-miR-6085, hsa-miR-6752-5pand hsa-miR-135a-3p, or a combination thereof, congeners thereof,transcripts thereof, or variants or derivatives thereof: and, optionallyin combination therewith, hsa-miR-1231, hsa-miR-1233-5p, hsa-miR-150-3p,hsa-miR-1225-3p, hsa-miR-92a-2-5p, hsa-miR-423-5p, hsa-miR-1268a,hsa-miR-128-2-5p and hsa-miR-24-3p or a combination thereof, congenersthereof, transcripts thereof, or variants or derivatives thereof: and,optionally in combination therewith, hsa-miR-4697-5p, hsa-miR-3197,hsa-miR-675-5p, hsa-miR-4486, hsa-miR-7107-5p, hsa-miR-23a-3p,hsa-miR-4667-5p, hsa-miR-451a, hsa-miR-3940-5p, hsa-miR-8059,hsa-miR-6813-5p, hsa-miR-4492, hsa-miR-4476 and hsa-miR-6090 or acombination thereof, congeners thereof, transcripts thereof, or variantsor derivatives thereof.

The expression level of each target nucleic acid described above isincreased or decreased (hereinafter, referred to as“increased/decreased”) according to the type of the target nucleic acidin a subject who has colorectal cancer as compared with a healthysubject. Hence, the nucleic acid of the present invention can beeffectively used for measuring the expression level of the targetnucleic acid described above in a body fluid derived from a subject(e.g., a human) suspected of having colorectal cancer and a body fluidderived from a healthy subject and detecting colorectal cancer by thecomparison thereof.

The nucleic acid probe or the primer that can be used in the presentinvention is a nucleic acid probe capable of specifically binding to apolynucleotide that consists of a nucleotide sequence represented by atleast one of SEQ ID NOs: 1 to 171 and 606 to 614, or a primer foramplifying a polynucleotide that consists of a nucleotide sequencerepresented by at least one of SEQ ID NOs: 1 to 171 and 606 to 614.

The nucleic acid probe or the primer that can be further used in thepresent invention can comprise a nucleic acid probe capable ofspecifically binding to a polynucleotide consisting of a nucleotidesequence represented by at least one of SEQ ID NOs: 172 to 180, or aprimer for amplifying a polynucleotide consisting of a nucleotidesequence represented by at least one of SEQ ID NOs: 172 to 180.

The nucleic acid probe or the primer that can be further used in thepresent invention can comprise a nucleic acid probe capable ofspecifically binding to a polynucleotide consisting of a nucleotidesequence represented by at least one of SEQ ID NOs: 181 to 194, or aprimer for amplifying a polynucleotide consisting of a nucleotidesequence represented by at least one of SEQ ID NOs: 181 to 194.

Specifically, these nucleic acid probes or primers comprise acombination of one or more polynucleotides selected from apolynucleotide group comprising nucleotide sequences represented by anyof SEQ ID NOs: 1 to 635 or nucleotide sequences derived from thenucleotide sequences by the replacement of u with t, and a complementarypolynucleotide group thereof, a polynucleotide group respectivelyhybridizing under stringent conditions (mentioned later) to DNAsconsisting of nucleotide sequences complementary to these nucleotidesequences, and a complementary polynucleotide group thereof, and apolynucleotide group comprising 15 or more, preferably 17 or moreconsecutive nucleotides in the nucleotide sequences of thesepolynucleotide groups. These polynucleotides can be used as nucleic acidprobes and primers for detecting the colorectal cancer markers as targetnucleic acids.

More specifically, examples of the nucleic acid probe or the primer thatcan be used in the present invention include one or morepolynucleotide(s) selected from the group consisting of the followingpolynucleotides (a) to (e):

(a) a polynucleotide consisting of a nucleotide sequence represented byany of SEQ ID NOs: 1 to 171 and 606 to 614 or a nucleotide sequencederived from the nucleotide sequence by the replacement of u with t, avariant thereof, a derivative thereof, or a fragment thereof comprising15 or more consecutive nucleotides,(b) a polynucleotide comprising a nucleotide sequence represented by anyof SEQ ID NOs: 1 to 171 and 606 to 614,(c) a polynucleotide consisting of a nucleotide sequence complementaryto a nucleotide sequence represented by any of SEQ ID NOs: 1 to 171 and606 to 614 or a nucleotide sequence derived from the nucleotide sequenceby the replacement of u with t, a variant thereof, a derivative thereof,or a fragment thereof comprising 15 or more consecutive nucleotides,(d) a polynucleotide comprising a nucleotide sequence complementary to anucleotide sequence represented by any of SEQ ID NOs: 1 to 171 and 606to 614 or a nucleotide sequence derived from the nucleotide sequence bythe replacement of u with t, and(e) a polynucleotide hybridizing under stringent conditions to any ofthe polynucleotides (a) to (d).

In addition to at least one or more polynucleotide(s) selected from thepolynucleotides (a) to (e), the nucleic acid probe or the primer thatcan be further used in the present invention can comprisepolynucleotides selected from the group consisting of the followingpolynucleotides (f) to (j):

(f) a polynucleotide consisting of a nucleotide sequence represented byany of SEQ ID NOs: 172 to 180 or a nucleotide sequence derived from thenucleotide sequence by the replacement of u with t, a variant thereof, aderivative thereof, or a fragment thereof comprising 15 or moreconsecutive nucleotides,(g) a polynucleotide comprising a nucleotide sequence represented by anyof SEQ ID NOs: 172 to 180,(h) a polynucleotide consisting of a nucleotide sequence complementaryto a nucleotide sequence represented by any of SEQ ID NOs: 172 to 180 ora nucleotide sequence derived from the nucleotide sequence by thereplacement of u with t, a variant thereof, a derivative thereof, or afragment thereof comprising 15 or more consecutive nucleotides,(i) a polynucleotide comprising a nucleotide sequence complementary to anucleotide sequence represented by any of SEQ ID NOs: 172 to 180 or anucleotide sequence derived from the nucleotide sequence by thereplacement of u with t, and(j) a polynucleotide hybridizing under stringent conditions to any ofthe polynucleotides (f) to (i).

In addition to at least one or more polynucleotide(s) selected from thepolynucleotides (a) to (j), the nucleic acid probe or the primer thatcan be further used in the present invention can comprisepolynucleotides selected from the group consisting of the followingpolynucleotides (k) to (o):

(k) a polynucleotide consisting of a nucleotide sequence represented byany of SEQ ID NOs: 181 to 194 or a nucleotide sequence derived from thenucleotide sequence by the replacement of u with t, a variant thereof, aderivative thereof, or a fragment thereof comprising 15 or moreconsecutive nucleotides,(l) a polynucleotide comprising a nucleotide sequence represented by anyof SEQ ID NOs:181 to 194,(m) a polynucleotide consisting of a nucleotide sequence complementaryto a nucleotide sequence represented by any of SEQ ID NOs: 181 to 194 ora nucleotide sequence derived from the nucleotide sequence by thereplacement of u with t, a variant thereof, a derivative thereof, or afragment thereof comprising 15 or more consecutive nucleotides,(n) a polynucleotide comprising a nucleotide sequence complementary to anucleotide sequence represented by any of SEQ ID NOs: 181 to 194 or anucleotide sequence derived from the nucleotide sequence by thereplacement of u with t, and(o) a polynucleotide hybridizing under stringent conditions to any ofthe polynucleotides (k) to (n).

For these polynucleotides, the “fragment thereof comprising 15 or moreconsecutive nucleotides” can contain the number of nucleotides in therange of, for example, from 15 consecutive nucleotides to less than thetotal number of nucleotides of the sequence, from 17 consecutivenucleotides to less than the total number of nucleotides of thesequence, or from 19 consecutive nucleotides to less than the totalnumber of nucleotides of the sequence, in the nucleotide sequence ofeach polynucleotide, though the fragment is not limited thereto.

These polynucleotides or the fragments thereof used in the presentinvention may each be DNA or may each be RNA.

The polynucleotides that can be used in the present invention can eachbe prepared by use of a general technique such as a DNA recombinationtechnique, PCR, or a method using an automatic DNA/RNA synthesizer.

The DNA recombination technique and the PCR can employ a techniquedescribed in, for example, Ausubel et al., Current Protocols inMolecular Biology, John Willey & Sons, US (1993); and Sambrook et al.,Molecular Cloning—A Laboratory Manual, Cold Spring Harbor LaboratoryPress, US (1989).

The human-derived hsa-miR-6726-5p, hsa-miR-4257, hsa-miR-6787-5p,hsa-miR-6780b-5p, hsa-miR-3131, hsa-miR-7108-5p, hsa-miR-1343-3p,hsa-miR-1247-3p, hsa-miR-4651, hsa-miR-6757-5p, hsa-miR-3679-5p,hsa-miR-7641, hsa-miR-6746-5p, hsa-miR-8072, hsa-miR-6741-5p,hsa-miR-1908-5p, hsa-miR-6857-5p, hsa-miR-4746-3p, hsa-miR-744-5p,hsa-miR-4792, hsa-miR-564, hsa-miR-6791-5p, hsa-miR-6825-5p,hsa-miR-6826-5p, hsa-miR-4665-3p, hsa-miR-4467, hsa-miR-3188,hsa-miR-6125, hsa-miR-6756-5p, hsa-miR-1228-3p, hsa-miR-8063,hsa-miR-8069, hsa-miR-6875-5p, hsa-miR-3185, hsa-miR-4433b-3p,hsa-miR-6887-5p, hsa-miR-128-1-5p, hsa-miR-6724-5p, hsa-miR-1914-3p,hsa-miR-1225-5p, hsa-miR-4419b, hsa-miR-7110-5p, hsa-miR-187-5p,hsa-miR-3184-5p, hsa-miR-204-3p, hsa-miR-5572, hsa-miR-6729-5p,hsa-miR-615-5p, hsa-miR-6749-5p, hsa-miR-6515-3p, hsa-miR-3937,hsa-miR-6840-3p, hsa-miR-6893-5p, hsa-miR-4728-5p, hsa-miR-6717-5p,hsa-miR-7113-3p, hsa-miR-4665-5p, hsa-miR-642b-3p, hsa-miR-7109-5p,hsa-miR-6842-5p, hsa-miR-4442, hsa-miR-4433-3p, hsa-miR-4707-5p,hsa-miR-6126, hsa-miR-4449, hsa-miR-4706, hsa-miR-1913, hsa-miR-602,hsa-miR-939-5p, hsa-miR-4695-5p, hsa-miR-711, hsa-miR-6816-5p,hsa-miR-4632-5p, hsa-miR-6721-5p, hsa-miR-7847-3p, hsa-miR-6132,hsa-miR-887-3p, hsa-miR-3679-3p, hsa-miR-6784-5p, hsa-miR-1249,hsa-miR-937-5p, hsa-miR-5195-3p, hsa-miR-6732-5p, hsa-miR-4417,hsa-miR-4281, hsa-miR-4734, hsa-miR-6766-3p, hsa-miR-663a, hsa-miR-4513,hsa-miR-6781-5p, hsa-miR-1227-5p, hsa-miR-6845-5p, hsa-miR-6798-5p,hsa-miR-3620-5p, hsa-miR-1915-5p, hsa-miR-4294, hsa-miR-642a-3p,hsa-miR-371a-5p, hsa-miR-940, hsa-miR-4450, hsa-miR-4723-5p,hsa-miR-1469, hsa-miR-6861-5p, hsa-miR-7975, hsa-miR-6879-5p,hsa-miR-6802-5p, hsa-miR-1268b, hsa-miR-663b, hsa-miR-125a-3p,hsa-miR-2861, hsa-miR-6088, hsa-miR-4758-5p, hsa-miR-296-3p,hsa-miR-6738-5p, hsa-miR-671-5p, hsa-miR-4454, hsa-miR-4516,hsa-miR-7845-5p, hsa-miR-4741, hsa-miR-92b-5p, hsa-miR-6795-5p,hsa-miR-6805-3p, hsa-miR-4725-3p, hsa-miR-6782-5p, hsa-miR-4688,hsa-miR-6850-5p, hsa-miR-6777-5p, hsa-miR-6785-5p, hsa-miR-7106-5p,hsa-miR-3663-3p, hsa-miR-6131, hsa-miR-1915-3p, hsa-miR-4532,hsa-miR-6820-5p, hsa-miR-4689, hsa-miR-4638-5p, hsa-miR-3656,hsa-miR-3621, hsa-miR-6769b-5p, hsa-miR-149-3p, hsa-miR-23b-3p,hsa-miR-3135b, hsa-miR-6848-5p, hsa-miR-6769a-5p, hsa-miR-4327,hsa-miR-6765-3p, hsa-miR-6716-5p, hsa-miR-6877-5p, hsa-miR-6727-5p,hsa-miR-4534, hsa-miR-614, hsa-miR-1202, hsa-miR-575, hsa-miR-6870-5p,hsa-miR-6722-3p, hsa-miR-7977, hsa-miR-4649-5p, hsa-miR-4675,hsa-miR-6075, hsa-miR-6779-5p, hsa-miR-4271, hsa-miR-3196,hsa-miR-6803-5p, hsa-miR-6789-5p, hsa-miR-4648, hsa-miR-4508,hsa-miR-4749-5p, hsa-miR-4505, hsa-miR-5698, hsa-miR-1199-5p,hsa-miR-4763-3p, hsa-miR-1231, hsa-miR-1233-5p, hsa-miR-150-3p,hsa-miR-1225-3p, hsa-miR-92a-2-5p, hsa-miR-423-5p, hsa-miR-1268a,hsa-miR-128-2-5p, hsa-miR-24-3p, hsa-miR-4697-5p, hsa-miR-3197,hsa-miR-675-5p, hsa-miR-4486, hsa-miR-7107-5p, hsa-miR-23a-3p,hsa-miR-4667-5p, hsa-miR-451a, hsa-miR-3940-5p, hsa-miR-8059,hsa-miR-6813-5p, hsa-miR-4492, hsa-miR-4476, hsa-miR-6090,hsa-miR-6836-3p, hsa-miR-3195, hsa-miR-718, hsa-miR-3178, hsa-miR-638,hsa-miR-4497, hsa-miR-6085, hsa-miR-6752-5p and hsa-miR-135a-3prepresented by SEQ ID NOs: 1 to 194 and 606 to 614 are known in the art,and their acquisition methods are also known as mentioned above.Therefore, each polynucleotide that can be used as a nucleic acid probeor a primer in the present invention can be prepared by cloning thegene.

Such a nucleic acid probe or a primer can be chemically synthesizedusing an automatic DNA synthesis apparatus. In general, aphosphoramidite method is used in this synthesis, and single-strandedDNA up to approximately 100 nucleotides can be automatically synthesizedby this method. The automatic DNA synthesis apparatus is commerciallyavailable from, for example, Polygen GmbH, ABI, or Applied Biosystems,Inc.

Alternatively, the polynucleotide of the present invention can also beprepared by a cDNA cloning method. The cDNA cloning technique canemploy, for example, microRNA Cloning Kit Wako.

In this context, the sequences of the nucleic acid probe and the primerfor detecting the polynucleotide consisting of a nucleotide sequencerepresented by any of SEQ ID NOs: 1 to 194 and 606 to 614 do not existas miRNAs or precursors thereof in vivo. For example, the nucleotidesequences represented by SEQ ID NO: 11 and SEQ ID NO: 78 are producedfrom the precursor represented by SEQ ID NO: 205. This precursor has ahairpin-like structure as shown in FIG. 1 , and the nucleotide sequencesrepresented by SEQ ID NO: 11 and SEQ ID NO: 78 have mismatch sequenceswith each other. Likewise, a nucleotide sequence completelycomplementary to the nucleotide sequence represented by SEQ ID NO: 11 orSEQ ID NO: 78 is not naturally produced in vivo. Therefore, the nucleicacid probe and the primer for detecting the nucleotide sequencerepresented by any of SEQ ID NOs: 1 to 194 and 606 to 614 each have anartificial nucleotide sequence that does not exist in vivo.

3. Kit or Device for Detection of Colorectal Cancer

The present invention also provides a kit or a device for the detectionof colorectal cancer, comprising one or more polynucleotide(s) (whichcan include a variant, a fragment, and a derivative; hereinafter, alsoreferred to as a polynucleotide for detection) that can be used as anucleic acid probe or a primer in the present invention for measuring atarget nucleic acid as a colorectal cancer marker.

The target nucleic acid as a colorectal cancer marker according to thepresent invention is preferably selected from the following group 1:

miR-6726-5p, miR-4257, miR-6787-5p, miR-6780b-5p, miR-3131, miR-7108-5p,miR-1343-3p, miR-1247-3p, miR-4651, miR-6757-5p, miR-3679-5p, miR-7641,miR-6746-5p, miR-8072, miR-6741-5p, miR-1908-5p, miR-6857-5p,miR-4746-3p, miR-744-5p, miR-4792, miR-564, miR-6791-5p, miR-6825-5p,miR-6826-5p, miR-4665-3p, miR-4467, miR-3188, miR-6125, miR-6756-5p,miR-1228-3p, miR-8063, miR-8069, miR-6875-5p, miR-3185, miR-4433b-3p,miR-6887-5p, miR-128-1-5p, miR-6724-5p, miR-1914-3p, miR-1225-5p,miR-4419b, miR-7110-5p, miR-187-5p, miR-3184-5p, miR-204-3p, miR-5572,miR-6729-5p, miR-615-5p, miR-6749-5p, miR-6515-3p, miR-3937,miR-6840-3p, miR-6893-5p, miR-4728-5p, miR-6717-5p, miR-7113-3p,miR-4665-5p, miR-642b-3p, miR-7109-5p, miR-6842-5p, miR-4442,miR-4433-3p, miR-4707-5p, miR-6126, miR-4449, miR-4706, miR-1913,miR-602, miR-939-5p, miR-4695-5p, miR-711, miR-6816-5p, miR-4632-5p,miR-6721-5p, miR-7847-3p, miR-6132, miR-887-3p, miR-3679-3p,miR-6784-5p, miR-1249, miR-937-5p, miR-5195-3p, miR-6732-5p, miR-4417,miR-4281, miR-4734, miR-6766-3p, miR-663a, miR-4513, miR-6781-5p,miR-1227-5p, miR-6845-5p, miR-6798-5p, miR-3620-5p, miR-1915-5p,miR-4294, miR-642a-3p, miR-371a-5p, miR-940, miR-4450, miR-4723-5p,miR-1469, miR-6861-5p, miR-7975, miR-6879-5p, miR-6802-5p, miR-1268b,miR-663b, miR-125a-3p, miR-2861, miR-6088, miR-4758-5p, miR-296-3p,miR-6738-5p, miR-671-5p, miR-4454, miR-4516, miR-7845-5p, miR-4741,miR-92b-5p, miR-6795-5p, miR-6805-3p, miR-4725-3p, miR-6782-5p,miR-4688, miR-6850-5p, miR-6777-5p, miR-6785-5p, miR-7106-5p,miR-3663-3p, miR-6131, miR-1915-3p, miR-4532, miR-6820-5p, miR-4689,miR-4638-5p, miR-3656, miR-3621, miR-6769b-5p, miR-149-3p, miR-23b-3p,miR-3135b, miR-6848-5p, miR-6769a-5p, miR-4327, miR-6765-3p,miR-6716-5p, miR-6877-5p, miR-6727-5p, miR-4534, miR-614, miR-1202,miR-575, miR-6870-5p, miR-6722-3p, miR-7977, miR-4649-5p, miR-4675,miR-6075, miR-6779-5p, miR-4271, miR-3196, miR-6803-5p, miR-6789-5p,miR-4648, miR-4508, miR-4749-5p, miR-4505, miR-5698, miR-1199-5p,miR-4763-3p, miR-6836-3p, miR-3195, miR-718, miR-3178, miR-638,miR-4497, miR-6085, miR-6752-5p and miR-135a-3p.

An additional target nucleic acid that can be optionally used in themeasurement is selected from the following group 2: miR-1231,miR-1233-5p, miR-150-3p, miR-1225-3p, miR-92a-2-5p, miR-423-5p,miR-1268a, miR-128-2-5p and miR-24-3p.

An additional target nucleic acid that can be optionally further used inthe measurement is selected from the following group 3: miR-4697-5p,miR-3197, miR-675-5p, miR-4486, miR-7107-5p, miR-23a-3p, miR-4667-5p,miR-451a, miR-3940-5p, miR-8059, miR-6813-5p, miR-4492, miR-4476, andmiR-6090.

The kit or the device of the present invention comprises one or morenucleic acid(s) capable of specifically binding to any of the targetnucleic acids as the colorectal cancer markers described above,preferably one or more polynucleotide(s) selected from the nucleic acidprobes or the primers described in the preceding Section 2,specifically, the polynucleotides described in the preceding Section 2,or variant(s) thereof.

Specifically, the kit or the device of the present invention cancomprise at least one or more polynucleotide(s) comprising (orconsisting of) a nucleotide sequence represented by any of SEQ ID NOs: 1to 171 and 606 to 614 or a nucleotide sequence derived from thenucleotide sequence by the replacement of u with t, polynucleotide(s)comprising (or consisting of) a complementary sequence thereof,polynucleotide(s) hybridizing under stringent conditions to any of thesepolynucleotides, or variant(s) or fragment(s) comprising 15 or moreconsecutive nucleotides of any of these polynucleotide sequences.

The kit or the device of the present invention can further comprise oneor more polynucleotide(s) comprising (or consisting of) a nucleotidesequence represented by any of SEQ ID NOs: 172 to 180 or a nucleotidesequence derived from the nucleotide sequence by the replacement of uwith t, polynucleotide(s) comprising (or consisting of) a complementarysequence thereof, polynucleotide(s) hybridizing under stringentconditions to any of these polynucleotides, variant(s) or fragment(s)comprising 15 or more consecutive nucleotides of any of thesepolynucleotide sequences.

The kit or the device of the present invention can further comprise oneor more polynucleotide(s) comprising (or consisting of) a nucleotidesequence represented by any of SEQ ID NOs: 181 to 194 or a nucleotidesequence derived from the nucleotide sequence by the replacement of uwith t, polynucleotide(s) comprising (or consisting of) a complementarysequence thereof, polynucleotide(s) hybridizing under stringentconditions to any of these polynucleotides, variant(s) or fragment(s)comprising 15 or more consecutive nucleotides of any of thesepolynucleotide sequences.

The fragment that can be contained in the kit or the device of thepresent invention is, for example, one or more, preferably two or morepolynucleotides selected from the group consisting of the followingpolynucleotides (1) to (3):

(1) a polynucleotide comprising 15 or more consecutive nucleotides in anucleotide sequence derived from a nucleotide sequence represented byany of SEQ ID NOs: 1 to 171 and 606 to 614 by the replacement of u witht, or a complementary sequence thereof;(2) a polynucleotide comprising 15 or more consecutive nucleotides in anucleotide sequence derived from a nucleotide sequence represented byany of SEQ ID NOs: 172 to 180 by the replacement of u with t, or acomplementary sequence thereof; and(3) a polynucleotide comprising 15 or more consecutive nucleotides in anucleotide sequence derived from a nucleotide sequence represented byany of SEQ ID NOs: 181 to 194 by the replacement of u with t, or acomplementary sequence thereof.

In a preferred embodiment, the polynucleotide is a polynucleotideconsisting of a nucleotide sequence represented by any of SEQ ID NOs: 1to 171 and 606 to 614 or a nucleotide sequence derived from thenucleotide sequence by the replacement of u with t, a polynucleotideconsisting of a complementary sequence thereof, a polynucleotidehybridizing under stringent conditions to any of these polynucleotides,or a variant thereof comprising 15 or more, preferably 17 or more, morepreferably 19 or more consecutive nucleotides.

In a preferred embodiment, the polynucleotide is a polynucleotideconsisting of a nucleotide sequence represented by any of SEQ ID NOs:172 to 180 or a nucleotide sequence derived from the nucleotide sequenceby the replacement of u with t, a polynucleotide consisting of acomplementary sequence thereof, a polynucleotide hybridizing understringent conditions to any of these polynucleotides, or a variantthereof comprising 15 or more, preferably 17 or more, more preferably 19or more consecutive nucleotides.

In a preferred embodiment, the polynucleotide is a polynucleotideconsisting of a nucleotide sequence represented by any of SEQ ID NOs:181 to 194 or a nucleotide sequence derived from the nucleotide sequenceby the replacement of u with t, a polynucleotide consisting of acomplementary sequence thereof, a polynucleotide hybridizing understringent conditions to any of these polynucleotides, or a variantthereof comprising 15 or more, preferably 17 or more, more preferably 19or more consecutive nucleotides.

In a preferred embodiment, the fragment can be a polynucleotidecomprising 15 or more, preferably 17 or more, more preferably 19 or moreconsecutive nucleotides.

In the present invention, the size of the polynucleotide fragment is thenumber of bases in the range of, for example, from 15 consecutivenucleotides to less than the total number of bases of the sequence, from17 consecutive nucleotides to less than the total number of bases of thesequence, or from 19 consecutive nucleotides to less than the totalnumber of nucleotides of the sequence, in the nucleotide sequence ofeach polynucleotide.

Specific examples of the aforementioned polynucleotide combinationconstituting the kit or the device of the present invention can includecombinations of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more of thepolynucleotides consisting of nucleotide sequences represented by SEQ IDNOs shown in Table 1 (SEQ ID NOs: 1 to 194 and 606 to 614 correspondingto the miRNA markers in the table). However, these are given merely forillustrative purposes, and various other possible combinations areincluded in the present invention.

The combination constituting the kit or the device for discriminating acolorectal cancer patient from a healthy subject according to thepresent invention is desirably, for example, a combination of two ormore of the polynucleotides consisting of nucleotide sequencesrepresented by SEQ ID NOs shown in Table 1. Usually, a combination oftwo of these polynucleotides can produce adequate performance.

The specific combination of two polynucleotides consisting of thenucleotide sequences or the complementary sequences thereof fordiscriminating a colorectal cancer patient from a healthy subject ispreferably a combination comprising at least one or more of newly foundpolynucleotides consisting of the nucleotide sequences represented bySEQ ID NOs: 1 to 171, among the combinations constituted by two of theaforementioned polynucleotides consisting of the nucleotide sequencesrepresented by SEQ ID NOs: 1 to 194 and 606 to 614. More specifically, acombination comprising at least one of polynucleotides consisting of thenucleotide sequences represented by SEQ ID NOs: 5, 15, 24, 32, 38, 45,55, 64, 96, 97, and 162, among the combinations of the polynucleotidesconsisting of the nucleotide sequences represented by SEQ ID NOs: 1 to194 and 606 to 614, is more preferred.

The combination of polynucleotides with cancer type specificity capableof discriminating a colorectal cancer patient not only from a healthysubject but also from other cancer patients is preferably, for example,a combination of multiple polynucleotides comprising at least onepolynucleotide selected from the group consisting of polynucleotides ofSEQ ID NOs: 5, 13, 15, 24, 32, 38, 41, 45, 55, 57, 64, 72, 75, 77, 96,97, 115, 162, 163, 173, 189, 606, 607, 608, 609, 610, 611, 612, 613 and614 (hereinafter, this group is referred to as “cancer type-specificpolynucleotide group 1”), with any of the polynucleotides of the otherSEQ ID NOs.

The combination of polynucleotides with cancer type specificity capableof discriminating a colorectal cancer patient not only from a healthysubject but also from other cancer patients is more preferably acombination of multiple polynucleotides selected from the cancertype-specific polynucleotide group 1.

The combination of polynucleotides with cancer type specificity capableof discriminating a colorectal cancer patient not only from a healthysubject but also from other cancer patients is more preferably acombination comprising at least one or more polynucleotide(s) selectedfrom the group consisting of polynucleotides of SEQ ID NOs: 5, 45, 57,96, and 606 (hereinafter, this group is referred to as “cancertype-specific polynucleotide group 2”) included in the cancertype-specific polynucleotide group 1, among the combinations of multiplepolynucleotides selected from the cancer type-specific polynucleotidegroup 1.

The number of the aforementioned polynucleotides with cancer typespecificity in the combination can be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 ormore for the combination and is more preferably 6 or more for thecombination. Usually, the combination of 5 or 6 of these polynucleotidescan produce adequate performance.

Non-limiting examples of the combination of the polynucleotideconsisting of the nucleotide sequence represented by SEQ ID NO: 5 or acomplementary sequence thereof with polynucleotides consisting ofnucleotide sequences represented by SEQ ID NOs of four or fivepolynucleotides selected from the cancer type-specific polynucleotidegroup 1 or complementary sequences thereof will be listed.

(1) a combination of SEQ ID NOs: 5, 45, 57, 75, and 607 (markers:hsa-miR-3131, hsa-miR-204-3p, hsa-miR-4665-5p, hsa-miR-7847-3p, andhsa-miR-3195);

(2) a combination of SEQ ID NOs: 5, 45, 96, 606, and 607 (markers:hsa-miR-3131, hsa-miR-204-3p, hsa-miR-4294, hsa-miR-6836-3p, andhsa-miR-3195);

(3) a combination of SEQ ID NOs: 5, 45, 57, 97, 115, and 607 (markers:hsa-miR-3131, hsa-miR-204-3p, hsa-miR-4665-5p, hsa-miR-642a-3p,hsa-miR-671-5p, and hsa-miR-3195);

(4) a combination of SEQ ID NOs: 5, 45, 57, 97, 162, and 607 (markers:hsa-miR-3131, hsa-miR-204-3p, hsa-miR-4665-5p, hsa-miR-642a-3p,hsa-miR-3196, and hsa-miR-3195);

(5) a combination of SEQ ID NOs: 5, 45, 57, 162, 607, and 613 (markers:hsa-miR-3131, hsa-miR-204-3p, hsa-miR-4665-5p, hsa-miR-3196,hsa-miR-3195, and hsa-miR-6752-5p);

(6) a combination of SEQ ID NOs: 5, 45, 57, 97, 607, and 612 (markers:hsa-miR-3131, hsa-miR-204-3p, hsa-miR-4665-5p, hsa-miR-642a-3p,hsa-miR-3195, and hsa-miR-6085);

(7) a combination of SEQ ID NOs: 5, 13, 45, 57, 606, and 607 (markers:hsa-miR-3131, hsa-miR-6746-5p, hsa-miR-204-3p, hsa-miR-4665-5p,hsa-miR-6836-3p, and hsa-miR-3195);

(8) a combination of SEQ ID NOs: 5, 45, 96, 189, 606, and 608 (markers:hsa-miR-3131, hsa-miR-204-3p, hsa-miR-4294, hsa-miR-3940-5p,hsa-miR-6836-3p, and hsa-miR-718);

(9) a combination of SEQ ID NOs: 5, 45, 57, 96, 189, and 606 (markers:hsa-miR-3131, hsa-miR-204-3p, hsa-miR-4665-5p, hsa-miR-4294,hsa-miR-3940-5p, and hsa-miR-6836-3p);

(10) a combination of SEQ ID NOs: 5, 24, 45, 57, 96, and 608 (markers:hsa-miR-3131, hsa-miR-6826-5p, hsa-miR-204-3p, hsa-miR-4665-5p,hsa-miR-4294, and hsa-miR-718);

(11) a combination of SEQ ID NOs: 5, 45, 57, 162, 607, and 610 (markers:hsa-miR-3131, hsa-miR-204-3p, hsa-miR-4665-5p, hsa-miR-3196,hsa-miR-3195, and hsa-miR-638); and

(12) a combination of SEQ ID NOs: 5, 45, 57, 189, 606, and 607 (markers:hsa-miR-3131, hsa-miR-204-3p, hsa-miR-4665-5p, hsa-miR-3940-5p,hsa-miR-6836-3p, and hsa-miR-3195).

Non-limiting examples of the combination of the polynucleotideconsisting of the nucleotide sequence represented by SEQ ID NO: 45 or acomplementary sequence thereof with polynucleotides consisting ofnucleotide sequences represented by SEQ ID NOs of four or fivepolynucleotides selected from the cancer type-specific polynucleotidegroup 1 or complementary sequences thereof will be further listed.

(1) a combination of SEQ ID NOs: 5, 45, 96, 606, and 607 (markers:hsa-miR-3131, hsa-miR-204-3p, hsa-miR-4294, hsa-miR-6836-3p, andhsa-miR-3195);

(2) a combination of SEQ ID NOs: 5, 45, 57, 75, and 607 (markers:hsa-miR-3131, hsa-miR-204-3p, hsa-miR-4665-5p, hsa-miR-7847-3p, andhsa-miR-3195);

(3) a combination of SEQ ID NOs: 5, 45, 57, 75, 606, and 607 (markers:hsa-miR-3131, hsa-miR-204-3p, hsa-miR-4665-5p, hsa-miR-7847-3p,hsa-miR-6836-3p, and hsa-miR-3195);

(4) a combination of SEQ ID NOs: 5, 45, 57, 77, 607, and 613 (markers:hsa-miR-3131, hsa-miR-204-3p, hsa-miR-4665-5p, hsa-miR-887-3p,hsa-miR-3195, and hsa-miR-6752-5p);

(5) a combination of SEQ ID NOs: 5, 45, 57, 97, 606, and 607 (markers:hsa-miR-3131, hsa-miR-204-3p, hsa-miR-4665-5p, hsa-miR-642a-3p,hsa-miR-6836-3p, and hsa-miR-3195);

(6) a combination of SEQ ID NOs: 5, 45, 57, 75, 77, and 607 (markers:hsa-miR-3131, hsa-miR-204-3p, hsa-miR-4665-5p, hsa-miR-7847-3p,hsa-miR-887-3p, and hsa-miR-3195);

(7) a combination of SEQ ID NOs: 5, 32, 45, 57, 96, and 606 (markers:hsa-miR-3131, hsa-miR-8069, hsa-miR-204-3p, hsa-miR-4665-5p,hsa-miR-4294, and hsa-miR-6836-3p);

(8) a combination of SEQ ID NOs: 5, 24, 45, 57, 96, and 606 (markers:hsa-miR-3131, hsa-miR-6826-5p, hsa-miR-204-3p, hsa-miR-4665-5p,hsa-miR-4294, and hsa-miR-6836-3p);

(9) a combination of SEQ ID NOs: 5, 45, 57, 96, 162, and 606 (markers:hsa-miR-3131, hsa-miR-204-3p, hsa-miR-4665-5p, hsa-miR-4294,hsa-miR-3196, and hsa-miR-6836-3p);

(10) a combination of SEQ ID NOs: 5, 15, 45, 75, 96, and 606 (markers:hsa-miR-3131, hsa-miR-6741-5p, hsa-miR-204-3p, hsa-miR-7847-3p,hsa-miR-4294, and hsa-miR-6836-3p);

(11) a combination of SEQ ID NOs: 5, 32, 45, 57, 162, and 607 (markers:hsa-miR-3131, hsa-miR-8069, hsa-miR-204-3p, hsa-miR-4665-5p,hsa-miR-3196, and hsa-miR-3195); and

(12) a combination of SEQ ID NOs: 38, 45, 96, 606, 608, and 611(markers: hsa-miR-6724-5p, hsa-miR-204-3p, hsa-miR-4294,hsa-miR-6836-3p, hsa-miR-718, and hsa-miR-4497).

Non-limiting examples of the combination of the polynucleotideconsisting of the nucleotide sequence represented by SEQ ID NO: 57 or acomplementary sequence thereof with polynucleotides consisting ofnucleotide sequences represented by SEQ ID NOs of four or fivepolynucleotides selected from the cancer type-specific polynucleotidegroup 1 or complementary sequences thereof will be further listed.

(1) a combination of SEQ ID NOs: 24, 41, 57, 45, and 96 (markers:hsa-miR-6826-5p, hsa-miR-4419b, hsa-miR-4665-5p, hsa-miR-204-3p, andhsa-miR-4294);

(2) a combination of SEQ ID NOs: 5, 45, 57, 607, and 612 (markers:hsa-miR-3131, hsa-miR-204-3p, hsa-miR-4665-5p, hsa-miR-3195, andhsa-miR-6085);

(3) a combination of SEQ ID NOs: 5, 45, 57, 606, 607, and 608 (markers:hsa-miR-3131, hsa-miR-204-3p, hsa-miR-4665-5p, hsa-miR-6836-3p,hsa-miR-3195, and hsa-miR-718);

(4) a combination of SEQ ID NOs: 5, 13, 45, 57, 75, and 607 (markers:hsa-miR-3131, hsa-miR-6746-5p, hsa-miR-204-3p, hsa-miR-4665-5p,hsa-miR-7847-3p, and hsa-miR-3195);

(5) a combination of SEQ ID NOs: 5, 45, 57, 64, 75, and 607 (markers:hsa-miR-3131, hsa-miR-204-3p, hsa-miR-4665-5p, hsa-miR-6126,hsa-miR-7847-3p, and hsa-miR-3195);

(6) a combination of SEQ ID NOs: 5, 45, 55, 57, 607, and 613 (markers:hsa-miR-3131, hsa-miR-204-3p, hsa-miR-6717-5p, hsa-miR-4665-5p,hsa-miR-3195, and hsa-miR-6752-5p);

(7) a combination of SEQ ID NOs: 5, 45, 55, 57, 75, and 607 (markers:hsa-miR-3131, hsa-miR-204-3p, hsa-miR-6717-5p, hsa-miR-4665-5p,hsa-miR-7847-3p, and hsa-miR-3195);

(8) a combination of SEQ ID NOs: 5, 38, 45, 57, 96, and 607 (markers:hsa-miR-3131, hsa-miR-6724-5p, hsa-miR-204-3p, hsa-miR-4665-5p,hsa-miR-4294, and hsa-miR-3195);

(9) a combination of SEQ ID NOs: 5, 45, 57, 75, 162, and 607 (markers:hsa-miR-3131, hsa-miR-204-3p, hsa-miR-4665-5p, hsa-miR-7847-3p,hsa-miR-3196, and hsa-miR-3195);

(10) a combination of SEQ ID NOs: 5, 45, 57, 75, 162, and 609 (markers:hsa-miR-3131, hsa-miR-204-3p, hsa-miR-4665-5p, hsa-miR-7847-3p,hsa-miR-3196, and hsa-miR-3178);

(11) a combination of SEQ ID NOs: 5, 45, 57, 64, 96, and 607 (markers:hsa-miR-3131, hsa-miR-204-3p, hsa-miR-4665-5p, hsa-miR-6126,hsa-miR-4294, and hsa-miR-3195); and

(12) a combination of SEQ ID NOs: 57, 64, 96, 606, 608, and 611(markers: hsa-miR-4665-5p, hsa-miR-6126, hsa-miR-4294, hsa-miR-6836-3p,hsa-miR-718, and hsa-miR-4497).

Non-limiting examples of the combination of the polynucleotideconsisting of the nucleotide sequence represented by SEQ ID NO: 96 or acomplementary sequence thereof with polynucleotides consisting ofnucleotide sequences represented by SEQ ID NOs of four or fivepolynucleotides selected from the cancer type-specific polynucleotidegroup 1 or complementary sequences thereof will be further listed below.

(1) a combination of SEQ ID NOs: 38, 96, 606, 608, and 611 (markers:hsa-miR-6724-5p, hsa-miR-4294, hsa-miR-6836-3p, hsa-miR-718, andhsa-miR-4497);

(2) a combination of SEQ ID NOs: 5, 45, 57, 96, and 607 (markers:hsa-miR-3131, hsa-miR-204-3p, hsa-miR-4665-5p, hsa-miR-4294, andhsa-miR-3195);

(3) a combination of SEQ ID NOs: 38, 72, 96, 606, 608, and 611 (markers:hsa-miR-6724-5p, hsa-miR-6816-5p, hsa-miR-4294, hsa-miR-6836-3p,hsa-miR-718, and hsa-miR-4497);

(4) a combination of SEQ ID NOs: 32, 38, 96, 606, 608, and 611 (markers:hsa-miR-8069, hsa-miR-6724-5p, hsa-miR-4294, hsa-miR-6836-3p,hsa-miR-718, and hsa-miR-4497);

(5) a combination of SEQ ID NOs: 38, 96, 163, 606, 608, and 611(markers: hsa-miR-6724-5p, hsa-miR-4294, hsa-miR-6803-5p,hsa-miR-6836-3p, hsa-miR-718, and hsa-miR-4497);

(6) a combination of SEQ ID NOs: 64, 72, 96, 162, 609, and 611 (markers:hsa-miR-6126, hsa-miR-6816-5p, hsa-miR-4294, hsa-miR-3196, hsa-miR-3178,and hsa-miR-4497);

(7) a combination of SEQ ID NOs: 38, 64, 96, 163, 606, and 608 (markers:hsa-miR-6724-5p, hsa-miR-6126, hsa-miR-4294, hsa-miR-6803-5p,hsa-miR-6836-3p, and hsa-miR-718);

(8) a combination of SEQ ID NOs: 5, 45, 57, 75, 96, and 606 (markers:hsa-miR-3131, hsa-miR-204-3p, hsa-miR-4665-5p, hsa-miR-7847-3p,hsa-miR-4294, and hsa-miR-6836-3p);

(9) a combination of SEQ ID NOs: 5, 15, 45, 57, 96, and 606 (markers:hsa-miR-3131, hsa-miR-6741-5p, hsa-miR-204-3p, hsa-miR-4665-5p,hsa-miR-4294, and hsa-miR-6836-3p);

(10) a combination of SEQ ID NOs: 5, 41, 45, 57, 96, and 606 (markers:hsa-miR-3131, hsa-miR-4419b, hsa-miR-204-3p, hsa-miR-4665-5p,hsa-miR-4294, and hsa-miR-6836-3p);

(11) a combination of SEQ ID NOs: 5, 41, 45, 96, 189, and 606 (markers:hsa-miR-3131, hsa-miR-4419b, hsa-miR-204-3p, hsa-miR-4294,hsa-miR-3940-5p, and hsa-miR-6836-3p); and

(12) a combination of SEQ ID NOs: 5, 45, 75, 96, 189, and 606 (markers:hsa-miR-3131, hsa-miR-204-3p, hsa-miR-7847-3p, hsa-miR-4294,hsa-miR-3940-5p, and hsa-miR-6836-3p).

Non-limiting examples of the combination of the polynucleotideconsisting of the nucleotide sequence represented by SEQ ID NO: 606 or acomplementary sequence thereof with polynucleotides consisting ofnucleotide sequences represented by SEQ ID NOs of three polynucleotidesselected from the cancer type-specific polynucleotide group 1 orcomplementary sequences thereof will be further listed.

(1) a combination of SEQ ID NOs: 5, 24, 45, 96, 189, and 606 (markers:hsa-miR-3131, hsa-miR-6826-5p, hsa-miR-204-3p, hsa-miR-4294,hsa-miR-3940-5p, and hsa-miR-6836-3p);

(2) a combination of SEQ ID NOs: 5, 15, 45, 96, 189, and 606 (markers:hsa-miR-3131, hsa-miR-6741-5p, hsa-miR-204-3p, hsa-miR-4294,hsa-miR-3940-5p, and hsa-miR-6836-3p);

(3) a combination of SEQ ID NOs: 5, 45, 96, 189, 606, and 613 (markers:hsa-miR-3131, hsa-miR-204-3p, hsa-miR-4294, hsa-miR-3940-5p,hsa-miR-6836-3p, and hsa-miR-6752-5p);

(4) a combination of SEQ ID NOs: 5, 45, 72, 96, 189, and 606 (markers:hsa-miR-3131, hsa-miR-204-3p, hsa-miR-6816-5p, hsa-miR-4294,hsa-miR-3940-5p, and hsa-miR-6836-3p); and

(5) a combination of SEQ ID NOs: 5, 15, 32, 45, 96, and 606 (markers:hsa-miR-3131, hsa-miR-6741-5p, hsa-miR-8069, hsa-miR-204-3p,hsa-miR-4294, and hsa-miR-6836-3p).

The kit or the device of the present invention can also contain apolynucleotide that is already known or that will be found in thefuture, to enable detection of colorectal cancer, in addition to thepolynucleotide(s) (which can include a variant, a fragment, and aderivative) according to the present invention described above.

The kit of the present invention can also contain an antibody formeasuring a marker for colorectal cancer examination known in the art,such as CEA or CA19-9, in addition to the polynucleotide(s) according tothe present invention described above.

These polynucleotides contained in the kit of the present invention canbe packaged in different containers either individually or in anycombination.

The kit of the present invention can contain a kit for extracting anucleic acid (e.g., total RNA) from body fluids, cells, or tissues, afluorescent material for labeling, an enzyme and a medium for nucleicacid amplification, an instruction manual, etc.

The device of the present invention is a device for cancer markermeasurement in which nucleic acids such as the polynucleotides accordingto the present invention described above are bonded or attached to, forexample, a solid phase. Examples of the material for the solid phaseinclude plastics, paper, glass, and silicon. The material for the solidphase is preferably a plastic from the viewpoint of easy processability.The solid phase has any shape and is, for example, square, round,reed-shaped, or film-shaped. The device of the present inventionincludes, for example, a device for measurement by a hybridizationtechnique. Specific examples thereof include blotting devices andnucleic acid arrays (e.g., microarrays, DNA chips, and RNA chips).

The nucleic acid array technique is a technique which involves bindingor attaching the nucleic acids one by one by use of a method [e.g., amethod of spotting the nucleic acids using a high-density dispensercalled spotter or arrayer onto the surface of the solid phasesurface-treated, if necessary, by coating with L-lysine or theintroduction of a functional group such as an amino group or a carboxylgroup, a method of spraying the nucleic acids onto the solid phase usingan inkjet which injects very small liquid droplets by a piezoelectricelement or the like from a nozzle, or a method of sequentiallysynthesizing nucleotides on the solid phase] to prepare an array such asa chip and measuring a target nucleic acid through the use ofhybridization using this array.

The kit or the device of the present invention comprises nucleic acidscapable of specifically binding to the polynucleotides of at least oneor more, preferably at least two or more, more preferably at least threeor more, most preferably at least five or more to all of the colorectalcancer marker miRNAs, respectively, of group 1 described above. The kitor the device of the present invention can optionally further comprisenucleic acids capable of specifically binding to the polynucleotides ofat least one or more, preferably at least two or more, more preferablyat least three or more, most preferably at least five or more to all ofthe colorectal cancer marker miRNAs, respectively, of group 2 describedabove. The kit or the device of the present invention can optionallyfurther comprise nucleic acids capable of specifically binding to thepolynucleotides of at least one or more, preferably at least two ormore, more preferably at least three or more, most preferably at leastfive or more to all of the colorectal cancer marker miRNAs,respectively, of group 3 described above.

The kit or the device of the present invention can be used for detectingcolorectal cancer as described in the Section 4 below.

4. Method for Detecting Colorectal Cancer

The present invention further provides a method for detecting colorectalcancer, comprising using the kit or the device of the present invention(including the nucleic acid(s) that can be used in the presentinvention) described in the preceding Section 3 above to measure anexpression level of one or more colorectal cancer-derived gene(s)represented by an expression level of colorectal cancer-derived gene(s)selected from the following group: miR-6726-5p, miR-4257, miR-6787-5p,miR-6780b-5p, miR-3131, miR-7108-5p, miR-1343-3p, miR-1247-3p, miR-4651,miR-6757-5p, miR-3679-5p, miR-7641, miR-6746-5p, miR-8072, miR-6741-5p,miR-1908-5p, miR-6857-5p, miR-4746-3p, miR-744-5p, miR-4792, miR-564,miR-6791-5p, miR-6825-5p, miR-6826-5p, miR-4665-3p, miR-4467, miR-3188,miR-6125, miR-6756-5p, miR-1228-3p, miR-8063, miR-8069, miR-6875-5p,miR-3185, miR-4433b-3p, miR-6887-5p, miR-128-1-5p, miR-6724-5p,miR-1914-3p, miR-1225-5p, miR-4419b, miR-7110-5p, miR-187-5p,miR-3184-5p, miR-204-3p, miR-5572, miR-6729-5p, miR-615-5p, miR-6749-5p,miR-6515-3p, miR-3937, miR-6840-3p, miR-6893-5p, miR-4728-5p,miR-6717-5p, miR-7113-3p, miR-4665-5p, miR-642b-3p, miR-7109-5p,miR-6842-5p, miR-4442, miR-4433-3p, miR-4707-5p, miR-6126, miR-4449,miR-4706, miR-1913, miR-602, miR-939-5p, miR-4695-5p, miR-711,miR-6816-5p, miR-4632-5p, miR-6721-5p, miR-7847-3p, miR-6132,miR-887-3p, miR-3679-3p, miR-6784-5p, miR-1249, miR-937-5p, miR-5195-3p,miR-6732-5p, miR-4417, miR-4281, miR-4734, miR-6766-3p, miR-663a,miR-4513, miR-6781-5p, miR-1227-5p, miR-6845-5p, miR-6798-5p,miR-3620-5p, miR-1915-5p, miR-4294, miR-642a-3p, miR-371a-5p, miR-940,miR-4450, miR-4723-5p, miR-1469, miR-6861-5p, miR-7975, miR-6879-5p,miR-6802-5p, miR-1268b, miR-663b, miR-125a-3p, miR-2861, miR-6088,miR-4758-5p, miR-296-3p, miR-6738-5p, miR-671-5p, miR-4454, miR-4516,miR-7845-5p, miR-4741, miR-92b-5p, miR-6795-5p, miR-6805-3p,miR-4725-3p, miR-6782-5p, miR-4688, miR-6850-5p, miR-6777-5p,miR-6785-5p, miR-7106-5p, miR-3663-3p, miR-6131, miR-1915-3p, miR-4532,miR-6820-5p, miR-4689, miR-4638-5p, miR-3656, miR-3621, miR-6769b-5p,miR-149-3p, miR-23b-3p, miR-3135b, miR-6848-5p, miR-6769a-5p, miR-4327,miR-6765-3p, miR-6716-5p, miR-6877-5p, miR-6727-5p, miR-4534, miR-614,miR-1202, miR-575, miR-6870-5p, miR-6722-3p, miR-7977, miR-4649-5p,miR-4675, miR-6075, miR-6779-5p, miR-4271, miR-3196, miR-6803-5p,miR-6789-5p, miR-4648, miR-4508, miR-4749-5p, miR-4505, miR-5698,miR-1199-5p, miR-4763-3p, miR-6836-3p, miR-3195, miR-718, miR-3178,miR-638, miR-4497, miR-6085, miR-6752-5p and miR-135a-3p, optionally anexpression level of colorectal cancer-derived gene(s) selected from thefollowing group: miR-1231, miR-1233-5p, miR-150-3p, miR-1225-3p,miR-92a-2-5p, miR-423-5p, miR-1268a, miR-128-2-5p and miR-24-3p, andoptionally an expression level of colorectal cancer-derived gene(s)selected from the following group: miR-4697-5p, miR-3197, miR-675-5p,miR-4486, miR-7107-5p, miR-23a-3p, miR-4667-5p, miR-451a, miR-3940-5p,miR-8059, miR-6813-5p, miR-4492, miR-4476, and miR-6090 in a sample invitro, further comparing, for example, the expression level of theaforementioned gene in the sample (e.g., blood, serum, or plasma)collected from a subject suspected of having colorectal cancer with acontrol expression level in the sample collected from a healthy subject(including a non-colorectal cancer patient), and evaluating the subjectas having colorectal cancer when the expression level of the targetnucleic acid is statistically significantly different between thesamples.

This method of the present invention permits lowly-invasive earlydiagnosis of cancer with high sensitivity and specificity and therebybrings about early treatment and improved prognosis. In addition,exacerbation of the disease or the effectiveness of surgical,radiotherapeutic, and chemotherapeutic treatments can be monitored.

The method for extracting the colorectal cancer-derived gene from thesample such as blood, serum, or plasma according to the presentinvention is particularly preferably prepared by the addition of areagent for RNA extraction in 3D-Gene™ RNA extraction reagent fromliquid sample kit (Toray Industries, Inc.). A general acidic phenolmethod (acid guanidinium-phenol-chloroform (AGPC)) may be used, orTrizol™ (Life Technologies Corp.) may be used. The colorectalcancer-derived gene may be prepared by the addition of a reagent for RNAextraction containing acidic phenol, such as Trizol (Life TechnologiesCorp.) or Isogen (Nippon Gene Co., Ltd.). Alternatively, a kit such asmiRNeasy™ Mini Kit (Qiagen N.V.) can be used, though the method is notlimited thereto.

The present invention also provides use of the kit or the device of thepresent invention for detecting in vitro an expression product of acolorectal cancer-derived miRNA gene in a sample derived from a subject.

In the method of the present invention, a kit or a device comprising,each alone or in every possible composition, the polynucleotides thatcan be used in the present invention as described above is used as thekit or the device.

In the detection or (genetic) diagnosis of colorectal cancer accordingto the present invention, each polynucleotide contained in the kit orthe device of the present invention can be used as a probe or a primer.In the case of using the polynucleotide as a primer, TaqMan™ MicroRNAAssays from Life Technologies Corp., miScript PCR System from QiagenN.V., or the like can be used, though the method is not limited thereto.

The polynucleotide contained in the kit or the device of the presentinvention can be used as a primer or a probe according to a routinemethod in a method known in the art for specifically detecting theparticular gene, for example, a hybridization technique such as Northernblot, Southern blot, in situ hybridization, Northern hybridization, orSouthern hybridization, or a quantitative amplification technique suchas quantitative RT-PCR. A body fluid such as blood, serum, plasma, orurine from a subject is collected as a sample to be assayed according tothe type of the detection method used. Alternatively, total RNA preparedfrom such a body fluid by the method described above may be used, andvarious polynucleotides including cDNA prepared on the basis of the RNAmay be used.

The kit or the device of the present invention is useful for thediagnosis of colorectal cancer or the detection of the presence orabsence of colorectal cancer. Specifically, the detection of colorectalcancer using the kit or the device can be performed by detecting invitro an expression level of a gene using the nucleic acid probe or theprimer contained in the kit or the device in a sample such as blood,serum, plasma, or urine from a subject suspected of having colorectalcancer. The subject suspected of having colorectal cancer can beevaluated as having colorectal cancer when the expression level of atarget miRNA marker measured using polynucleotide(s) (including avariant, a fragment, and a derivative thereof) consisting of anucleotide sequence represented by at least one or more of SEQ ID NOs: 1to 171 and 606 to 614 or a complementary sequence thereof, optionally anucleotide sequence represented by one or more of SEQ ID NOs: 172 to 180or a complementary sequence thereof, and optionally a nucleotidesequence represented by one or more of SEQ ID NOs: 181 to 194 or acomplementary sequence thereof in the sample such as blood, serum,plasma, or urine of the subject is statistically significantly differentfrom the expression level thereof in the sample such as blood, serum, orplasma, or urine of a healthy subject.

The method of the present invention can be combined with fecal occultblood, rectal examination, and colonoscopy as well as a diagnosticimaging method such as barium enema, CT, MRI, or bone scintigraphy. Themethod of the present invention is capable of specifically detectingcolorectal cancer and can substantially discriminate colorectal cancerfrom other cancers.

The method for detecting the absence of an expression product of acolorectal cancer-derived gene or the presence of the expression productof a colorectal cancer-derived gene in a sample using the kit or thedevice of the present invention comprises; collecting a body fluid suchas blood, serum, plasma, or urine from a subject, measuring theexpression level of the target gene that contains therein using one ormore polynucleotide(s) (including a variant, a fragment, and aderivative) selected from the polynucleotide group of the presentinvention, and evaluating the presence or absence of colorectal canceror detecting colorectal cancer. Using the method for detectingcolorectal cancer according to the present invention, for example, thepresence or absence of amelioration of the disease or the degree ofamelioration thereof in a colorectal cancer patient given a therapeuticdrug for the amelioration of the disease can be also evaluated ordiagnosed.

The method of the present invention can comprise, for example, thefollowing steps (a), (b), and (c):

(a) a step of contacting a sample derived from a subject with apolynucleotide in the kit or the device of the present invention invitro;

(b) a step of measuring an expression level of the target nucleic acidin the sample using the polynucleotide as a nucleic acid probe or aprimer; and

(c) a step of evaluating the presence or absence of colorectal cancer(cells) in the subject on the basis of the step (b).

Specifically, the present invention provides a method for detectingcolorectal cancer, comprising measuring an expression level of a targetnucleic acid in a sample of a subject using a nucleic acid capable ofspecifically binding to at least one or more (preferably at least two ormore) polynucleotide(s) selected from the group consisting ofmiR-6726-5p, miR-4257, miR-6787-5p, miR-6780b-5p, miR-3131, miR-7108-5p,miR-1343-3p, miR-1247-3p, miR-4651, miR-6757-5p, miR-3679-5p, miR-7641,miR-6746-5p, miR-8072, miR-6741-5p, miR-1908-5p, miR-6857-5p,miR-4746-3p, miR-744-5p, miR-4792, miR-564, miR-6791-5p, miR-6825-5p,miR-6826-5p, miR-4665-3p, miR-4467, miR-3188, miR-6125, miR-6756-5p,miR-1228-3p, miR-8063, miR-8069, miR-6875-5p, miR-3185, miR-4433b-3p,miR-6887-5p, miR-128-1-5p, miR-6724-5p, miR-1914-3p, miR-1225-5p,miR-4419b, miR-7110-5p, miR-187-5p, miR-3184-5p, miR-204-3p, miR-5572,miR-6729-5p, miR-615-5p, miR-6749-5p, miR-6515-3p, miR-3937,miR-6840-3p, miR-6893-5p, miR-4728-5p, miR-6717-5p, miR-7113-3p,miR-4665-5p, miR-642b-3p, miR-7109-5p, miR-6842-5p, miR-4442,miR-4433-3p, miR-4707-5p, miR-6126, miR-4449, miR-4706, miR-1913,miR-602, miR-939-5p, miR-4695-5p, miR-711, miR-6816-5p, miR-4632-5p,miR-6721-5p, miR-7847-3p, miR-6132, miR-887-3p, miR-3679-3p,miR-6784-5p, miR-1249, miR-937-5p, miR-5195-3p, miR-6732-5p, miR-4417,miR-4281, miR-4734, miR-6766-3p, miR-663a, miR-4513, miR-6781-5p,miR-1227-5p, miR-6845-5p, miR-6798-5p, miR-3620-5p, miR-1915-5p,miR-4294, miR-642a-3p, miR-371a-5p, miR-940, miR-4450, miR-4723-5p,miR-1469, miR-6861-5p, miR-7975, miR-6879-5p, miR-6802-5p, miR-1268b,miR-663b, miR-125a-3p, miR-2861, miR-6088, miR-4758-5p, miR-296-3p,miR-6738-5p, miR-671-5p, miR-4454, miR-4516, miR-7845-5p, miR-4741,miR-92b-5p, miR-6795-5p, miR-6805-3p, miR-4725-3p, miR-6782-5p,miR-4688, miR-6850-5p, miR-6777-5p, miR-6785-5p, miR-7106-5p,miR-3663-3p, miR-6131, miR-1915-3p, miR-4532, miR-6820-5p, miR-4689,miR-4638-5p, miR-3656, miR-3621, miR-6769b-5p, miR-149-3p, miR-23b-3p,miR-3135b, miR-6848-5p, miR-6769a-5p, miR-4327, miR-6765-3p,miR-6716-5p, miR-6877-5p, miR-6727-5p, miR-4534, miR-614, miR-1202,miR-575, miR-6870-5p, miR-6722-3p, miR-7977, miR-4649-5p, miR-4675,miR-6075, miR-6779-5p, miR-4271, miR-3196, miR-6803-5p, miR-6789-5p,miR-4648, miR-4508, miR-4749-5p, miR-4505, miR-5698, miR-1199-5p andmiR-4763-3p, miR-6836-3p, miR-3195, miR-718, miR-3178, miR-638,miR-4497, miR-6085, miR-6752-5p and miR-135a-3p and evaluating in vitrowhether or not the subject has colorectal cancer using the measuredexpression level and a control expression level of a healthy subjectmeasured in the same way as above.

As used herein, the term “evaluation” is evaluation support based onresults of in vitro examination, not physician's judgment.

As described above, as for the target nucleic acids in a preferredembodiment of the method of the present invention, specifically,miR-6726-5p is hsa-miR-6726-5p, miR-4257 is hsa-miR-4257, miR-6787-5p ishsa-miR-6787-5p, miR-6780b-5p is hsa-miR-6780b-5p, miR-3131 ishsa-miR-3131, miR-7108-5p is hsa-miR-7108-5p, miR-1343-3p ishsa-miR-1343-3p, miR-1247-3p is hsa-miR-1247-3p, miR-4651 ishsa-miR-4651, miR-6757-5p is hsa-miR-6757-5p, miR-3679-5p ishsa-miR-3679-5p, miR-7641 is hsa-miR-7641, miR-6746-5p ishsa-miR-6746-5p, miR-8072 is hsa-miR-8072, miR-6741-5p ishsa-miR-6741-5p, miR-1908-5p is hsa-miR-1908-5p, miR-6857-5p ishsa-miR-6857-5p, miR-4746-3p is hsa-miR-4746-3p, miR-744-5p ishsa-miR-744-5p, miR-4792 is hsa-miR-4792, miR-564 is hsa-miR-564,miR-6791-5p is hsa-miR-6791-5p, miR-6825-5p is hsa-miR-6825-5p,miR-6826-5p is hsa-miR-6826-5p, miR-4665-3p is hsa-miR-4665-3p, miR-4467is hsa-miR-4467, miR-3188 is hsa-miR-3188, miR-6125 is hsa-miR-6125,miR-6756-5p is hsa-miR-6756-5p, miR-1228-3p is hsa-miR-1228-3p, miR-8063is hsa-miR-8063, miR-8069 is hsa-miR-8069, miR-6875-5p ishsa-miR-6875-5p, miR-3185 is hsa-miR-3185, miR-4433b-3p ishsa-miR-4433b-3p, miR-6887-5p is hsa-miR-6887-5p, miR-128-1-5p ishsa-miR-128-1-5p, miR-6724-5p is hsa-miR-6724-5p, miR-1914-3p ishsa-miR-1914-3p, miR-1225-5p is hsa-miR-1225-5p, miR-4419b ishsa-miR-4419b, miR-7110-5p is hsa-miR-7110-5p, miR-187-5p ishsa-miR-187-5p, miR-3184-5p is hsa-miR-3184-5p, miR-204-3p ishsa-miR-204-3p, miR-5572 is hsa-miR-5572, miR-6729-5p ishsa-miR-6729-5p, miR-615-5p is hsa-miR-615-5p, miR-6749-5p ishsa-miR-6749-5p, miR-6515-3p is hsa-miR-6515-3p, miR-3937 ishsa-miR-3937, miR-6840-3p is hsa-miR-6840-3p, miR-6893-5p ishsa-miR-6893-5p, miR-4728-5p is hsa-miR-4728-5p, miR-6717-5p ishsa-miR-6717-5p, miR-7113-3p is hsa-miR-7113-3p, miR-4665-5p ishsa-miR-4665-5p, miR-642b-3p is hsa-miR-642b-3p, miR-7109-5p ishsa-miR-7109-5p, miR-6842-5p is hsa-miR-6842-5p, miR-4442 ishsa-miR-4442, miR-4433-3p is hsa-miR-4433-3p, miR-4707-5p ishsa-miR-4707-5p, miR-6126 is hsa-miR-6126, miR-4449 is hsa-miR-4449,miR-4706 is hsa-miR-4706, miR-1913 is hsa-miR-1913, miR-602 ishsa-miR-602, miR-939-5p is hsa-miR-939-5p, miR-4695-5p ishsa-miR-4695-5p, miR-711 is hsa-miR-711, miR-6816-5p is hsa-miR-6816-5p,miR-4632-5p is hsa-miR-4632-5p, miR-6721-5p is hsa-miR-6721-5p,miR-7847-3p is hsa-miR-7847-3p, miR-6132 is hsa-miR-6132, miR-887-3p ishsa-miR-887-3p, miR-3679-3p is hsa-miR-3679-3p, miR-6784-5p ishsa-miR-6784-5p, miR-1249 is hsa-miR-1249, miR-937-5p is hsa-miR-937-5p,miR-5195-3p is hsa-miR-5195-3p, miR-6732-5p is hsa-miR-6732-5p, miR-4417is hsa-miR-4417, miR-4281 is hsa-miR-4281, miR-4734 is hsa-miR-4734,miR-6766-3p is hsa-miR-6766-3p, miR-663a is hsa-miR-663a, miR-4513 ishsa-miR-4513, miR-6781-5p is hsa-miR-6781-5p, miR-1227-5p ishsa-miR-1227-5p, miR-6845-5p is hsa-miR-6845-5p, miR-6798-5p ishsa-miR-6798-5p, miR-3620-5p is hsa-miR-3620-5p, miR-1915-5p ishsa-miR-1915-5p, miR-4294 is hsa-miR-4294, miR-642a-3p ishsa-miR-642a-3p, miR-371a-5p is hsa-miR-371a-5p, miR-940 is hsa-miR-940,miR-4450 is hsa-miR-4450, miR-4723-5p is hsa-miR-4723-5p, miR-1469 ishsa-miR-1469, miR-6861-5p is hsa-miR-6861-5p, miR-7975 is hsa-miR-7975,miR-6879-5p is hsa-miR-6879-5p, miR-6802-5p is hsa-miR-6802-5p,miR-1268b is hsa-miR-1268b, miR-663b is hsa-miR-663b, miR-125a-3p ishsa-miR-125a-3p, miR-2861 is hsa-miR-2861, miR-6088 is hsa-miR-6088,miR-4758-5p is hsa-miR-4758-5p, miR-296-3p is hsa-miR-296-3p,miR-6738-5p is hsa-miR-6738-5p, miR-671-5p is hsa-miR-671-5p, miR-4454is hsa-miR-4454, miR-4516 is hsa-miR-4516, miR-7845-5p ishsa-miR-7845-5p, miR-4741 is hsa-miR-4741, miR-92b-5p is hsa-miR-92b-5p,miR-6795-5p is hsa-miR-6795-5p, miR-6805-3p is hsa-miR-6805-3p,miR-4725-3p is hsa-miR-4725-3p, miR-6782-5p is hsa-miR-6782-5p, miR-4688is hsa-miR-4688, miR-6850-5p is hsa-miR-6850-5p, miR-6777-5p ishsa-miR-6777-5p, miR-6785-5p is hsa-miR-6785-5p, miR-7106-5p ishsa-miR-7106-5p, miR-3663-3p is hsa-miR-3663-3p, miR-6131 ishsa-miR-6131, miR-1915-3p is hsa-miR-1915-3p, miR-4532 is hsa-miR-4532,miR-6820-5p is hsa-miR-6820-5p, miR-4689 is hsa-miR-4689, miR-4638-5p ishsa-miR-4638-5p, miR-3656 is hsa-miR-3656, miR-3621 is hsa-miR-3621,miR-6769b-5p is hsa-miR-6769b-5p, miR-149-3p is hsa-miR-149-3p,miR-23b-3p is hsa-miR-23b-3p, miR-3135b is hsa-miR-3135b, miR-6848-5p ishsa-miR-6848-5p, miR-6769a-5p is hsa-miR-6769a-5p, miR-4327 ishsa-miR-4327, miR-6765-3p is hsa-miR-6765-3p, miR-6716-5p ishsa-miR-6716-5p, miR-6877-5p is hsa-miR-6877-5p, miR-6727-5p ishsa-miR-6727-5p, miR-4534 is hsa-miR-4534, miR-614 is hsa-miR-614,miR-1202 is hsa-miR-1202, miR-575 is hsa-miR-575, miR-6870-5p ishsa-miR-6870-5p, miR-6722-3p is hsa-miR-6722-3p, miR-7977 ishsa-miR-7977, miR-4649-5p is hsa-miR-4649-5p, miR-4675 is hsa-miR-4675,miR-6075 is hsa-miR-6075, miR-6779-5p is hsa-miR-6779-5p, miR-4271 ishsa-miR-4271, miR-3196 is hsa-miR-3196, miR-6803-5p is hsa-miR-6803-5p,miR-6789-5p is hsa-miR-6789-5p, miR-4648 is hsa-miR-4648, miR-4508 ishsa-miR-4508, miR-4749-5p is hsa-miR-4749-5p, miR-4505 is hsa-miR-4505,miR-5698 is hsa-miR-5698, miR-1199-5p is hsa-miR-1199-5p, miR-4763-3p ishsa-miR-4763-3p, miR-6836-3p is hsa-miR-6836-3p, miR-3195 ishsa-miR-3195, miR-718 is hsa-miR-718, miR-3178 is hsa-miR-3178, miR-638is hsa-miR-638, miR-4497 is hsa-miR-4497, miR-6085 is hsa-miR-6085,miR-6752-5p is hsa-miR-6752-5p, and miR-135a-3p is hsa-miR-135a-3p.

In a preferred embodiment of the method of the present invention,specifically, the nucleic acid (specifically, probe or primer) isselected from the group consisting of the following polynucleotides (a)to (e):

(a) a polynucleotide consisting of a nucleotide sequence represented byany of SEQ ID NOs: 1 to 171 and 606 to 614 or a nucleotide sequencederived from the nucleotide sequence by the replacement of u with t, avariant thereof, a derivative thereof, or a fragment thereof comprising15 or more consecutive nucleotides,(b) a polynucleotide comprising a nucleotide sequence represented by anyof SEQ ID NOs: 1 to 171 and 606 to 614,(c) a polynucleotide consisting of a nucleotide sequence complementaryto a nucleotide sequence represented by any of SEQ ID NOs: 1 to 171 and606 to 614 or a nucleotide sequence derived from the nucleotide sequenceby the replacement of u with t, a variant thereof, a derivative thereof,or a fragment thereof comprising 15 or more consecutive nucleotides,(d) a polynucleotide comprising a nucleotide sequence complementary to anucleotide sequence represented by any of SEQ ID NOs: 1 to 171 and 606to 614 or a nucleotide sequence derived from the nucleotide sequence bythe replacement of u with t, and(e) a polynucleotide hybridizing under stringent conditions to any ofthe polynucleotides (a) to (d).

The method of the present invention can further employ a nucleic acidcapable of specifically binding to at least one or morepolynucleotide(s) selected from the group consisting of miR-1231,miR-1233-5p, miR-150-3p, miR-1225-3p, miR-92a-2-5p, miR-423-5p,miR-1268a, miR-128-2-5p and miR-24-3p.

As for such a nucleic acid, specifically, miR-1231 is hsa-miR-1231,miR-1233-5p is hsa-miR-1233-5p, miR-150-3p is hsa-miR-150-3p,miR-1225-3p is hsa-miR-1225-3p, miR-92a-2-5p is hsa-miR-92a-2-5p,miR-423-5p is hsa-miR-423-5p, miR-1268a is hsa-miR-1268a, miR-128-2-5pis hsa-miR-128-2-5p, and miR-24-3p is hsa-miR-24-3p.

In a preferred embodiment, such a nucleic acid is specifically selectedfrom the group consisting of the following polynucleotides (f) to (j):

(f) a polynucleotide consisting of a nucleotide sequence represented byany of SEQ ID NOs: 172 to 180 or a nucleotide sequence derived from thenucleotide sequence by the replacement of u with t, a variant thereof, aderivative thereof, or a fragment thereof comprising 15 or moreconsecutive nucleotides,(g) a polynucleotide comprising a nucleotide sequence represented by anyof SEQ ID NOs: 172 to 180,(h) a polynucleotide consisting of a nucleotide sequence complementaryto a nucleotide sequence represented by any of SEQ ID NOs: 172 to 180 ora nucleotide sequence derived from the nucleotide sequence by thereplacement of u with t, a variant thereof, a derivative thereof, or afragment thereof comprising 15 or more consecutive nucleotides,(i) a polynucleotide comprising a nucleotide sequence complementary to anucleotide sequence represented by any of SEQ ID NOs: 172 to 180 or anucleotide sequence derived from the nucleotide sequence by thereplacement of u with t, and(j) a polynucleotide hybridizing under stringent conditions to any ofthe polynucleotides (f) to (i).

The nucleic acid further used in the method of the present invention cancomprise a nucleic acid capable of specifically binding to at least oneor more polynucleotide(s) selected from the group consisting ofmiR-4697-5p, miR-3197, miR-675-5p, miR-4486, miR-7107-5p, miR-23a-3p,miR-4667-5p, miR-451a, miR-3940-5p, miR-8059, miR-6813-5p, miR-4492,miR-4476 and miR-6090.

As for such a nucleic acid, specifically, miR-4697-5p ishsa-miR-4697-5p, miR-3197 is hsa-miR-3197, miR-675-5p is hsa-miR-675-5p,miR-4486 is hsa-miR-4486, miR-7107-5p is hsa-miR-7107-5p, miR-23a-3p ishsa-miR-23a-3p, miR-4667-5p is hsa-miR-4667-5p, miR-451a ishsa-miR-451a, miR-3940-5p is hsa-miR-3940-5p, miR-8059 is hsa-miR-8059,miR-6813-5p is hsa-miR-6813-5p, miR-4492 is hsa-miR-4492, miR-4476 ishsa-miR-4476, and miR-6090 is hsa-miR-6090.

In a preferred embodiment, such a nucleic acid is specifically apolynucleotide selected from the group consisting of the followingpolynucleotides (k) to (o):

(k) a polynucleotide consisting of a nucleotide sequence represented byany of SEQ ID NOs: 181 to 194 or a nucleotide sequence derived from thenucleotide sequence by the replacement of u with t, a variant thereof, aderivative thereof, or a fragment thereof comprising 15 or moreconsecutive nucleotides,(l) a polynucleotide comprising a nucleotide sequence represented by anyof SEQ ID NOs: 181 to 194,(m) a polynucleotide consisting of a nucleotide sequence complementaryto a nucleotide sequence represented by any of SEQ ID NOs: 181 to 194 ora nucleotide sequence derived from the nucleotide sequence by thereplacement of u with t, a variant thereof, a derivative thereof, or afragment thereof comprising 15 or more consecutive nucleotides,(n) a polynucleotide comprising a nucleotide sequence complementary to anucleotide sequence represented by any of SEQ ID NOs: 181 to 194 or anucleotide sequence derived from the nucleotide sequence by thereplacement of u with t, and(o) a polynucleotide hybridizing under stringent conditions to any ofthe polynucleotides (k) to (n).

Examples of the sample used in the method of the present invention caninclude samples prepared from a living tissue (preferably a colorectaltissue) or a body fluid such as blood, serum, plasma, or urine from thesubject. Specifically, for example, an RNA-containing sample preparedfrom the tissue, a polynucleotide-containing sample further preparedtherefrom, a body fluid such as blood, serum, plasma, or urine, aportion or the whole of a living tissue collected from the subject bybiopsy or the like, or a living tissue excised by surgery can be used,and the sample for measurement can be prepared therefrom.

As used herein, the subject refers to a mammal, for example, a human, amonkey, a mouse and a rat without any limitation, and is preferably ahuman.

The steps of the method of the present invention can be changedaccording to the type of the sample to be assayed.

In the case of using RNA as an analyte, the detection of colorectalcancer (cells) can comprise, for example, the following steps (a), (b),and (c):

(a) a step of binding RNA prepared from the sample of the subject or acomplementary polynucleotide (cDNA) transcribed therefrom to apolynucleotide in the kit or the device of the present invention;

(b) a step of measuring the sample-derived RNA or the cDNA synthesizedfrom the RNA, bound with the polynucleotide by hybridization using thepolynucleotide as a nucleic acid probe or by quantitative RT-PCR usingthe polynucleotide as a primer; and

(c) a step of evaluating the presence or absence of colorectal cancer(or colorectal cancer-derived gene expression) on the basis of themeasurement results of the step (b).

For example, various hybridization methods can be used for detecting,examining, evaluating, or diagnosing colorectal cancer (or colorectalcancer-derived gene expression) in vitro according to the presentinvention. For example, Northern blot, Southern blot, RT-PCR, DNA chipanalysis, in situ hybridization, Northern hybridization, or Southernhybridization can be used as such a hybridization method.

In the case of using the Northern blot, the presence or absence ofexpression of each gene or the expression level thereof in the RNA canbe detected or measured by use of the nucleic acid probe that can beused in the present invention. Specific examples thereof can include amethod which involves labeling the nucleic acid probe (or itscomplementary strand) with a radioisotope (³²P, ³³P, ³⁵S, etc.), afluorescent material, or the like, that hybridizes the labeled productwith the living tissue-derived RNA from a subject transferred to a nylonmembrane or the like according to a routine method, and then detectingand measuring a signal derived from the label (radioisotope orfluorescent material) on the formed DNA/RNA duplex using a radiationdetector (examples thereof can include BAS-1800 II (Fujifilm Corp.)) ora fluorescence detector (examples thereof can include STORM 865 (GEHealthcare Japan Corp.)).

In the case of using the quantitative RT-PCR, the presence or absence ofexpression of each gene or the expression level thereof in the RNA canbe detected or measured by use of the primer that can be used in thepresent invention. Specific examples thereof can include a method whichinvolves; preparing cDNA from the living tissue-derived RNA of a subjectaccording to a routine method, hybridizing a pair of primers (consistingof a plus strand and a reverse strand binding to the cDNA) of thepresent invention with the cDNA such that the region of each target genecan be amplified with the cDNA as a template, and performing PCRaccording to a routine method to detect the obtained double-strandedDNA. The method for detecting the double-stranded DNA can include amethod of performing the PCR using the primers labeled in advance with aradioisotope or a fluorescent material, a method of electrophoresing thePCR product on an agarose gel and staining the double-stranded DNA withethidium bromide or the like for detection, and a method of transferringthe produced double-stranded DNA to a nylon membrane or the likeaccording to a routine method and hybridizing the double-stranded DNA toa labeled nucleic acid probe for detection.

In the case of using the nucleic acid array analysis, an RNA chip or aDNA chip in which the nucleic acid probes (single-stranded ordouble-stranded) of the present invention is attached to a substrate(solid phase) is used. Regions that have the attached nucleic acidprobes are referred to as probe spots, and regions that have no attachednucleic acid probe are referred to as blank spots. A gene groupimmobilized on a solid-phase substrate is generally called a nucleicacid chip, a nucleic acid array, a microarray, or the like. The DNA orRNA array includes a DNA or RNA macroarray and a DNA or RNA microarray.The term “chip” used herein includes all of these arrays. 3D-Gene™ HumanmiRNA Oligo chip (Toray Industries, Inc.) can be used as the DNA chip,though the DNA chip is not limited thereto.

Examples of the measurement using the DNA chip can include, but are notlimited to, a method of detecting and measuring a signal derived fromthe label on the nucleic acid probes using an image detector (examplesthereof can include Typhoon 9410 (GE Healthcare Japan Corp.) and3D-Gene™ scanner (Toray Industries, Inc.)).

The “stringent conditions” used herein are, as mentioned above,conditions under which a nucleic acid probe hybridizes to its targetsequence to a larger extent (e.g., a measurement value equal to orlarger than a mean of background measurement values+a standard deviationof the background measurement values×2) than that for other sequences.

The stringent conditions are defined by hybridization and subsequentwashing conditions. The hybridization conditions involve, for example,but not limited to, 30° C. to 60° C. for 1 to 24 hours in a solutioncontaining SSC, a surfactant, formamide, dextran sulfate, a blockingagent, etc. In this context, 1×SSC is an aqueous solution (pH 7.0)containing 150 mM sodium chloride and 15 mM sodium citrate. Thesurfactant includes, for example, SDS (sodium dodecyl sulfate), Triton,or Tween. The hybridization conditions more preferably involve 3 to10×SSC and 0.1 to 1% SDS. Examples of the conditions of the washing,following the hybridization, which is another condition to define thestringent conditions, can include conditions involving continuouswashing at 30° C. in a solution containing 0.5×SSC and 0.1% SDS, at 30°C. in a solution containing 0.2×SSC and 0.1% SDS, and at 30° C. in a0.05×SSC solution. It is desirable that the complementary strand shouldmaintain its hybridized state with a target plus strand even by thewashing under such conditions. Specifically, examples of such acomplementary strand can include a strand consisting of a nucleotidesequence in a completely complementary relationship with the nucleotidesequence of the target plus strand, and a strand consisting of anucleotide sequence having at least 80%, preferably at least 85%, morepreferably at least 90% or at least 95%, for example, at least 98% or atleast 99% identity to the strand.

Other examples of the “stringent conditions” for the hybridization aredescribed in, for example, Sambrook, J. & Russel, D., Molecular Cloning,A LABORATORY MANUAL, Cold Spring Harbor Laboratory Press, published onJan. 15, 2001, Vol. 1, 7.42 to 7.45 and Vol. 2, 8.9 to 8.17, and can beused in the present invention.

Examples of the conditions for carrying out PCR using a polynucleotidefragment in the kit of the present invention as a primer includetreatment for approximately 15 seconds to 1 minute at 5 to 10° C. plus aTm value calculated from the sequence of the primer, using a PCR bufferhaving composition such as 10 mM Tris-HCL (pH 8.3), 50 mM KCL, and 1 to2 mM MgCl₂. Examples of the method for calculating such a Tm valueinclude Tm value=2×(the number of adenine residues+the number of thymineresidues)+4×(the number of guanine residues+the number of cytosineresidues).

In the case of using the quantitative RT-PCR, a commercially availablekit for measurement specially designed for quantitatively measuringmiRNA, such as TaqMan™ MicroRNA Assays (Life Technologies Corp.);LNA™-based MicroRNA PCR (Exiqon); or Ncode™ miRNA qRT-PCT kit(Invitrogen Corp.) may be used.

For the calculation of gene expression levels, statistical analysisdescribed in, for example, Statistical analysis of gene expressionmicroarray data (Speed T., Chapman and Hall/CRC), and A beginner's guideMicroarray gene expression data analysis (Causton H. C. et al.,Blackwell publishing) can be used in the present invention, though thecalculation method is not limited thereto. For example, twice,preferably 3 times, more preferably 6 times the standard deviation ofthe measurement values of the blank spots are added to the averagemeasurement value of the blank spots on the DNA chip, and probe spotshaving a signal value equal to or larger than the resulting value can beregarded as detection spots. Alternatively, the average measurementvalue of the blank spots is regarded as a background and can besubtracted from the measurement values of the probe spots to determinegene expression levels. A missing value for a gene expression level canbe excluded from the analyte, preferably replaced with the smallestvalue of the gene expression level in each DNA chip, or more preferablyreplaced with a value obtained by subtracting 0.1 from a logarithmicvalue of the smallest value of the gene expression level. In order toeliminate low-signal genes, only a gene having a gene expression levelof 2⁶, preferably 2⁸, more preferably 2¹⁰ or larger, in 20% or more,preferably 50% or more, more preferably 80% or more of the number ofmeasurement samples can be selected as the analyte. Examples of thenormalization of the gene expression level include, but are not limitedto, global normalization and quantile normalization (Bolstad, B. M. etal., 2003, Bioinformatics, Vol. 19, p. 185-193).

The present invention also provides a method comprising measuring atarget gene or gene expression level in a sample derived from a subjectusing the polynucleotide, the kit, or the device (e.g., chip) fordetection of the present invention, or a combination thereof, preparinga discriminant (discriminant function) with gene expression levels in asample derived from a colorectal cancer patient and a sample derivedfrom a healthy subject as supervising samples, and determining orevaluating the presence and/or absence of the colorectal cancer-derivedgene in the sample.

Specifically, the present invention further provides the methodcomprising: a first step of measuring in vitro an expression level of atarget gene (target nucleic acid) in multiple samples known to determineor evaluate the presence and/or absence of the colorectal cancer-derivedgene in the samples, using the polynucleotide, the kit, or the device(e.g., chip) for detection of the present invention, or a combinationthereof; a second step of preparing a discriminant with the measurementvalues of the expression level of the target gene obtained in the firststep as supervising samples; a third step of measuring in vitro anexpression level of the target gene in a sample derived from a subjectin the same way as in the first step; and a fourth step of assigning themeasurement value of the expression level of the target gene obtained inthe third step into the discriminant obtained in the second step, anddetermining or evaluating the presence or absence of the colorectalcancer-derived gene in the sample on the basis of the results obtainedfrom the discriminant, wherein the target gene can be detected using thepolynucleotide or using a polynucleotide for detection contained in thekit or the device (e.g., chip). In this context, the discriminant can beprepared by use of Fisher's discriminant analysis, nonlineardiscriminant analysis based on Mahalanobis' distance, neural network,Support Vector Machine (SVM), or the like, though the method is notlimited thereto.

When a clustering boundary is a straight line or a hyperplane, thelinear discriminant analysis is a method for determining the associationof a cluster using Formula 1 as a discriminant. In Formula 1, xrepresents an explanatory variable, w represents a coefficient of theexplanatory variable, and w0 represents a constant term.

$\begin{matrix}{{f(x)} = {w_{0} + {\sum\limits_{i = 1}^{f}{w_{i}x_{i}}}}} & {{Formula}1}\end{matrix}$

Values obtained from the discriminant are referred to as discriminantscores. The measurement values of a newly offered data set can beassigned as explanatory variables to the discriminant to determineclusters by the signs of the discriminant scores.

The Fisher's discriminant analysis, one type of linear discriminantanalysis, is a dimensionality reduction method for selecting a dimensionsuitable for discriminating classes, and constructs synthetic variablewith highly discriminant performance by focusing on the variance ofsynthetic variables and minimizing the variance of data having the samelabel (Venables, W. N. et al., Modern Applied Statistics with S. Fourthedition. Springer., 2002). In the Fisher's discriminant analysis,direction w of projection is determined so as to maximize Formula 2. Inthis Formula, μ represents an average input, ng represents the number ofdata associated to class g, and μg represents an average input of thedata associated to class g. The numerator and the denominator are theinter-classe variance and the intra-classe variance, respectively, wheneach data is projected in the direction of the vector w. Discriminantcoefficient wi is determined by maximizing this ratio (Takafumi Kanamoriet al., “Pattern Recognition”, Kyoritsu Shuppan Co., Ltd. (2009); andRichard O. et al., Pattern Classification Second Edition.,Wiley-Interscience, 2000).

$\begin{matrix}{{J(w)} = \frac{\sum\limits_{g = 1}^{G}{{n_{g}\left( {{w^{T}\mu_{g}} - {w^{T}\mu}} \right)}\left( {{w^{T}\mu_{g}} - {w^{T}\mu}} \right)^{T}}}{\sum\limits_{g = 1}^{G}{\sum\limits_{{i:y_{i}} = g}{\left( {{w^{T}x_{i}} - {w^{T}\mu_{g}}} \right)\left( {{w^{T}x_{i}} - {w^{T}\mu_{g}}} \right)}}}} & {{Formula}2}\end{matrix}$ subjectto $\begin{matrix}{{\mu = {\sum\limits_{i = 1}^{n}\frac{x_{i}}{n}}},} & {\mu_{g} = {\sum\limits_{{i:u_{i}} = g}^{n}\frac{x_{i}}{n_{g}}}}\end{matrix}$

The Mahalanobis' distance is calculated according to Formula 3 inconsideration of data correlation and can be used as nonlineardiscriminant analysis for determining a cluster having a closerMahalanobis' distance from each cluster as an associated cluster. Inthis Formula 3, μ represents a central vector of each cluster, and S-1represents an inverse matrix of the variance-covariance matrix of thecluster. The central vector is calculated from explanatory variable x,and an average vector, a median value vector, or the like can be used.

$\begin{matrix}{{D\left( {x,\mu} \right)} = \left\{ {\left( {x - \mu} \right)^{t}{S^{- 1}\left( {x - \mu} \right)}} \right\}^{\frac{1}{2}}} & {{Formula}3}\end{matrix}$

SVM is a discriminant analysis method devised by V. Vapnik (The Natureof Statistical Leaning Theory, Springer, 1995). Particular data pointsof a data set having known classes are defined as explanatory variables,and classes are defined as objective variables. A boundary plane calledhyperplane for correctly classifying the data set into the known classesis determined, and a discriminant for data classification is determinedusing the boundary plane. Then, the measurement values of a newlyoffered data set can be assigned as explanatory variables to thediscriminant to determine classes. In this respect, the result of thediscriminant analysis may be classes, may be a probability of beingclassified into correct classes, or may be the distance from thehyperplane. In SVM, a method of nonlinearly converting a feature vectorto a high dimension and performing linear discriminant analysis in thespace is known as a method for tackling nonlinear problems. Anexpression in which an inner product of two factors in a nonlinearlymapped space is expressed only by inputs in their original spaces iscalled kernel. Examples of the kernel can include a linear kernel, a RBF(Radial Basis Function) kernel, and a Gaussian kernel. While highlydimensional mapping is performed according to the kernel, the optimumdiscriminant, i.e., a discriminant, can be actually constructed by merecalculation according to the kernel, which avoids calculating featuresin the mapped space (e.g., Hideki Aso et al., Frontier of StatisticalScience 6 “Statistics of pattern recognition and learning—New conceptsand approaches”, Iwanami Shoten, Publishers (2004); Nello Cristianini etal., Introduction to SVM, Kyoritsu Shuppan Co., Ltd. (2008)).

C-support vector classification (C-SVC), one type of SVM, involvespreparing a hyperplane by supervising with the explanatory variables oftwo groups and classifying an unknown data set into either of the groups(C. Cortes et al., 1995, Machine Learning, Vol. 20, p. 273-297).

Exemplary calculation of the C-SVC discriminant that can be used in themethod of the present invention will be given below. First, all subjectsare divided into two groups, i.e., a colorectal cancer patient group anda healthy subject group. For example, colorectal tissue examination canbe used for each subject to be confirmed either as a colorectal cancerpatient or as a healthy subject.

Next, a data set consisting of comprehensive gene expression levels ofserum-derived samples of the two divided groups (hereinafter, this dataset is referred to as a training cohort) is prepared, and a C-SVCdiscriminant is determined by using explanatory variables that are genesfound to differ clearly in their gene expression levels between the twogroups, and objective variables (e.g., −1 and +1) that are the grouping.An optimizing objective function is represented by Formula 4 wherein erepresents all input vectors, y represents an objective variable, arepresents a Lagrange's undetermined multiplier vector, Q represents apositive definite matrix, and C represents a parameter for adjustingconstrained conditions.

$\begin{matrix}\begin{matrix}{{\underset{a}{\min}\frac{1}{2}a^{T}Qa} - {e^{T}a}} \\{\begin{matrix}{{subject}{to}} & {{{y^{T}a} = 0},} & {{0 \leq a_{i} \leq C},} & {{i = 1},\ldots,l,}\end{matrix}}\end{matrix} & {{Formula}4}\end{matrix}$

Formula 5 is a finally obtained discriminant, and an associated groupcan be determined on the basis of the sign of a value obtained accordingto the discriminant. In this Formula, x represents a support vector, yrepresents a label indicating the association of a group, a representsthe corresponding coefficient, b represents a constant term, and Krepresents a kernel function.

$\begin{matrix}{{f(x)} = {{sgn}\left( {{\overset{l}{\sum\limits_{i = 1}}{y_{i}a_{i}{K\left( {x_{i},x} \right)}}} + b} \right)}} & {{Formula}5}\end{matrix}$

For example, a RBF kernel defined by Formula 6 can be used as the kernelfunction. In this Formula, x represents a support vector, and yrepresents a kernel parameter for adjusting the complexity of thehyperplane.

K(x _(i) ,x _(j))=exp(−r∥x _(i) −x _(j)∥²)r<0  Formula 6

In addition, an approach such as neural network, k-nearest neighboralgorithms, decision trees, or logistic regression analysis can beselected as a method for determining or evaluating the presence and/orabsence of expression of a colorectal cancer-derived target gene in asample derived from a subject, or for evaluating the expression levelthereof by comparison with a control derived from a healthy subject.

The method of the present invention can comprise, for example, thefollowing steps (a), (b), and (c):

(a) a step of measuring an expression level of a target gene in tissuescontaining colorectal cancer-derived genes derived from colorectalcancer patients and/or samples already known to be tissues containing nocolorectal cancer-derived gene derived from healthy subjects, using thepolynucleotide, the kit, or the device (e.g., DNA chip) for detectionaccording to the present invention;

(b) a step of preparing the discriminants of Formulas 1 to 3, 5, and 6described above from the measurement values of the expression levelmeasured in the step (a); and

(c) measuring an expression level of the target gene in a sample derivedfrom a subject using the polynucleotide, the kit, or the device (e.g.,DNA chip) for detection according to the present invention, assigningthe obtained measurement value to the discriminants prepared in the step(b), and determining or evaluating the presence and/or absence of thecolorectal cancer-derived target gene in the sample, or evaluating theexpression level thereof by comparison with a healthy subject-derivedcontrol, on the basis of the obtained results. In this context, in thediscriminants of Formulas 1 to 3, 5, and 6, x represents an explanatoryvariable and includes a value obtained by measuring a polynucleotideselected from the polynucleotides described in Section 2 above, or afragment thereof, etc. Specifically, the explanatory variable fordiscriminating a colorectal cancer patient from a healthy subjectaccording to the present invention is a gene expression level selectedfrom, for example, the following expression levels (1) to (3):

(1) a gene expression level in the serum of a colorectal cancer patientor a healthy subject measured by any DNA comprising 15 or moreconsecutive nucleotides in a nucleotide sequence represented by any ofSEQ ID NOs: 1 to 171 and 606 to 614 or a complementary sequence thereof,

(2) a gene expression level in the serum of a colorectal cancer patientor a healthy subject measured by any DNA comprising 15 or moreconsecutive nucleotides in a nucleotide sequence represented by any ofSEQ ID NOs: 172 to 180 or a complementary sequence thereof, and

(3) a gene expression level in the serum of a colorectal cancer patientor a healthy subject measured by any DNA comprising 15 or moreconsecutive nucleotides in a nucleotide sequence represented by any ofSEQ ID NOs: 181 to 194 or a complementary sequence thereof.

As described above, for the method for determining or evaluating thepresence and/or absence of a colorectal cancer-derived gene in a samplederived from a subject, the preparation of a discriminant requires adiscriminant constructed from a training cohort. For enhancing thediscriminant accuracy of the discriminant, it is necessary for thediscriminant to use genes that show clear difference between two groupsin the training cohort.

Each gene that is used for an explanatory variable in a discriminant ispreferably determined as follows. First, comprehensive gene expressionlevels of a colorectal cancer patient group and comprehensive geneexpression levels of a healthy subject group in a training cohort areused as a data set, the degree of difference in the expression level ofeach gene between the two groups is determined through the use of, forexample, the P value of t test, which is parametric analysis, or the Pvalue of Mann-Whitney's U test or Wilcoxon test, which is nonparametricanalysis.

The gene can be regarded as being statistically significant when thecritical rate (significance level) of the P value obtained by the testis smaller than, for example, 5%, 1%, or 0.01%.

In order to correct an increased probability of type I error attributedto the repetition of a test, a method known in the art, for example,Bonfenoni or Holm method, can be used for the correction (e.g., YasushiNagata et al., “Basics of statistical multiple comparison methods”,Scientist Press Co., Ltd. (2007)). As an example of the Bonferronicorrection, for example, the P value obtained by a test is multiplied bythe number of repetitions of the test, i.e., the number of genes used inthe analysis, and the obtained value can be compared with a desiredsignificance level to suppress a probability of causing type I error inthe whole test.

Instead of the statistical test, the absolute value (fold change) of anexpression ratio of a median value of each gene expression level betweengene expression levels of a colorectal cancer patient group and geneexpression levels of a healthy subject group may be calculated to selecta gene that is used for an explanatory variable in a discriminant.Alternatively, ROC curves may be prepared using gene expression levelsof a colorectal cancer patient group and a healthy subject group, and agene that is used for an explanatory variable in a discriminant can beselected on the basis of an AUROC value.

Next, a discriminant that can be calculated by various methods describedabove is constructed using any number of genes that show largedifference in their gene expression levels determined here. Examples ofthe method for constructing a discriminant that produces the largestdiscriminant accuracy include a method of constructing a discriminant inevery combination of genes that satisfy the significance level of Pvalue, and a method of repetitively evaluating the genes for use in theconstruction of a discriminant while increasing the number of genes oneby one in a descending order of the difference in gene expression level(Furey T S. et al., 2000, Bioinformatics., Vol. 16, p. 906-14). A geneexpression level of another independent colorectal cancer patient orhealthy subject is assigned as an explanatory variable to thisdiscriminant, and a result of the discriminant analysis regarding thegroup to which this independent colorectal cancer patient or healthysubject associated, is calculated. Specifically, the found gene set fordiagnosis and the discriminant constructed using the gene set fordiagnosis can be evaluated in an independent sample group to find a moreuniversal gene set for diagnosis capable of detecting colorectal cancerand a more universal method for discriminating colorectal cancer.

Split-sample method is preferably used for evaluating the discriminantperformance (generality) of the discriminant. Specifically, a data setis divided into a training cohort and a validation cohort, and geneselection by a statistical test and discriminant construction areperformed in the training cohort. Accuracy, sensitivity, and specificityare calculated using results of discriminating a validation cohortaccording to the discriminant and a true group to which the validationcohort associated, to evaluate the discriminant performance. On theother hand, instead of dividing a data set, gene selection by astatistical test and discriminant preparation may be performed using allof samples, and accuracy, sensitivity, and specificity can be calculatedby the discriminant of newly prepared samples according to thediscriminant to evaluate the discriminant performance.

The present invention provides a polynucleotide for detection or fordisease diagnosis useful in the diagnosis and treatment of colorectalcancer, a method for detecting colorectal cancer using thepolynucleotide, and a kit and a device for the detection of colorectalcancer, comprising the polynucleotide. Particularly, in order to selecta gene for diagnosis and prepare a discriminant so as to exhibitaccuracy beyond a colorectal cancer diagnosis method using existingtumor markers CEA, a gene set for diagnosis and a discriminant for themethod of the present invention can be constructed, which exhibitaccuracy beyond CEA, for example, by comparing genes expressed in serumderived from a patient confirmed to be negative using CEA but finallyfound to have colorectal cancer by detailed examination such as computedtomography using a contrast medium, with genes expressed in serumderived from a patient who has no colorectal cancer.

For example, the gene set for diagnosis is set to any combinationselected from one or two or more of the polynucleotides based on anucleotide sequence represented by any of SEQ ID NOs: 1 to 171 and 606to 614 or a complementary sequence thereof as described above,optionally one or two or more of the polynucleotides based on anucleotide sequence represented by any of SEQ ID NOs: 172 to 180 or acomplementary sequence thereof, and optionally one or two or more of thepolynucleotides based on a nucleotide sequence represented by any of SEQID NOs: 181 to 194 or a complementary sequence thereof. Further, adiscriminant is constructed using expression levels of the gene set fordiagnosis in samples derived from class I colorectal cancer patients andsamples derived from class II healthy subjects as a result of tissuediagnosis. As a result, the presence or absence of colorectalcancer-derived genes in an unknown sample can be determined with 100%accuracy at the maximum by measuring expression levels of the gene setfor diagnosis in the unknown sample.

EXAMPLES

Hereinafter, the present invention will be described furtherspecifically with reference to Examples below. However, the scope of thepresent invention is not intended to be limited by these Examples.

Reference Example 1

<Collection of Samples from Colorectal Cancer Patients and HealthySubjects>

Serum was collected using VENOJECT II vacuum blood collecting tubeVP-AS109K60 (Terumo Corp.) from each of 100 healthy subjects and 34colorectal cancer patients (15 cases with stage I, 6 cases with stageIIA, 4 cases with stage IIIA, 6 cases with stage IIIB, 2 cases withstage IIIC, and 1 case with stage IV) who were confirmed to have noprimary cancer other than colorectal cancer after acquisition ofinformed consent, and used as a training cohort. Likewise, serum wascollected using VENOJECT II vacuum blood collecting tube VP-AS109K60(Terumo Corp.) from each of 50 healthy subjects and 16 colorectal cancerpatients (3 cases with stage I, 4 cases with stage IIA, 1 case withstage IIB, 2 cases with stage IIIB, 2 cases with stage IIIC, and 4 caseswith stage IV) who were confirmed to have no primary cancer other thancolorectal cancer after acquisition of informed consent, and used as avalidation cohort.

<Extraction of Total RNA>

Total RNA was obtained from 300 μL of the serum sample obtained fromeach of 200 persons in total of 150 healthy subjects and 50 colorectalcancer patients included in the training cohort and the validationcohort, using a reagent for RNA extraction in 3D-Gene™ RNA extractionreagent from liquid sample kit (Toray Industries, Inc.) according to theprotocol provided by the manufacturer.

<Measurement of Gene Expression Level>

miRNAs in the total RNA obtained from the serum sample of each of 200persons in total of 150 healthy subjects and 50 colorectal cancerpatients included in the training cohort and the validation cohort werefluorescently labeled using 3D-Gene™ miRNA Labeling kit (TorayIndustries, Inc.) according to the protocol (ver 2.20) provided by themanufacturer. The oligo DNA chip used was 3D-Gene™ Human miRNA Oligochip (Toray Industries, Inc.) with attached probes having sequencescomplementary to 2,555 miRNAs among the miRNAs registered in miRBaseRelease 20. Hybridization between the miRNAs in the total RNA and theprobes on the DNA chip under stringent conditions and washing followingthe hybridization were performed according to the protocol provided bythe manufacturer. The DNA chip was scanned using 3D-Gene™ scanner (TorayIndustries, Inc.) to obtain images. Fluorescence intensity was digitizedusing 3D-Gene™ Extraction (Toray Industries, Inc.). The digitizedfluorescence intensity was converted to a logarithmic value with a baseof 2 and used as a gene expression level, from which a blank value wassubtracted. A missing value was replaced with a value obtained bysubtracting 0.1 from a logarithmic value of the smallest value of thegene expression level in each DNA chip. As a result, the comprehensivegene expression levels of the miRNAs in the serum were obtained for the50 colorectal cancer patients and the 150 healthy subjects. Calculationand statistical analysis using the digitized gene expression levels ofthe miRNAs were carried out using R language 3.0.2 (R Development CoreTeam (2013). R: A language and environment for statistical computing. RFoundation for Statistical Computing, URL http://www.R-project.org/.)and MASS package 7.3-30 (Venables, W. N. & Ripley, B. D. (2002) ModernApplied Statistics with S. Fourth Edition. Springer, New York. ISBN0-387-95457-0).

Reference Example 2

<Collection of Samples from Patients with Cancer Other than ColorectalCancer>

Serum was collected using VENOJECT II vacuum blood collecting tubeVP-AS109K60 (Terumo Corp.) from each of 69 pancreatic cancer patients,66 biliary tract cancer patients, 30 stomach cancer patients, 33esophageal cancer patients, 32 liver cancer patients, and 15 benignpancreaticobiliary disease patients who were confirmed to have no cancerin other organs after acquisition of informed consent, and used as atraining cohort together with the samples of 34 colorectal cancerpatients and 103 healthy subjects of Reference Example 1. Likewise,serum was collected using VENOJECT II vacuum blood collecting tubeVP-AS109K60 (Terumo Corp.) from each of 30 pancreatic cancer patients,33 bile duct cancer patients, 20 stomach cancer patients, 17 esophagealcancer patients, 20 liver cancer patients, and 6 benignpancreaticobiliary disease patients who were confirmed to have no cancerin other organs after acquisition of informed consent, and used as avalidation cohort together with the samples of 16 colorectal cancerpatients confirmed to have no cancer in organs other than the largeintestine and 47 healthy subjects of Reference Example 1. Subsequentoperations were conducted in the same way as in Reference Example 1.

Example 1

<Selection of Gene Marker Using Samples in the Training Cohort, andMethod for Evaluating Colorectal Cancer Discriminant Performance ofSingle Gene Marker Using Samples of in the Validation Cohort>

In this Example, a gene marker for discriminating a colorectal cancerpatient from a healthy subject was selected in the training cohort andstudied in samples in the validation cohort independent of the trainingcohort, for a method for evaluating the colorectal cancer discriminantperformance of each selected gene marker alone.

Specifically, first, the miRNA expression levels of the training cohortand the validation cohort obtained in the preceding Reference Exampleswere combined and normalized by quantile normalization.

Next, genes for diagnosis were selected using the training cohort. Here,in order to acquire diagnostic markers with higher reliability, onlygenes that show a gene expression level of 2⁶ or higher in 50% or moreof the samples in either of the colorectal cancer patient group in thetraining cohort or the healthy subject group in the training cohort wereselected. In order to further acquire statistically significant genesfor discriminating a colorectal cancer patient group from a healthysubject group, the P value obtained by two-tailed t-test assuming equalvariance as to each gene expression level was corrected by theBonferroni method, and genes that satisfied p<0.01 were acquired as genemarkers for use in explanatory variables of a discriminant. The obtainedgenes are described in Table 2.

In this way, hsa-miR-6726-5p, hsa-miR-4257, hsa-miR-6787-5p,hsa-miR-6780b-5p, hsa-miR-3131, hsa-miR-7108-5p, hsa-miR-1343-3p,hsa-miR-1247-3p, hsa-miR-4651, hsa-miR-6757-5p, hsa-miR-3679-5p,hsa-miR-7641, hsa-miR-6746-5p, hsa-miR-8072, hsa-miR-6741-5p,hsa-miR-1908-5p, hsa-miR-6857-5p, hsa-miR-4746-3p, hsa-miR-744-5p,hsa-miR-4792, hsa-miR-564, hsa-miR-6791-5p, hsa-miR-6825-5p,hsa-miR-6826-5p, hsa-miR-4665-3p, hsa-miR-4467, hsa-miR-3188,hsa-miR-6125, hsa-miR-6756-5p, hsa-miR-1228-3p, hsa-miR-8063,hsa-miR-8069, hsa-miR-6875-5p, hsa-miR-3185, hsa-miR-4433b-3p,hsa-miR-6887-5p, hsa-miR-128-1-5p, hsa-miR-6724-5p, hsa-miR-1914-3p,hsa-miR-1225-5p, hsa-miR-4419b, hsa-miR-7110-5p, hsa-miR-187-5p,hsa-miR-3184-5p, hsa-miR-204-3p, hsa-miR-5572, hsa-miR-6729-5p,hsa-miR-615-5p, hsa-miR-6749-5p, hsa-miR-6515-3p, hsa-miR-3937,hsa-miR-6840-3p, hsa-miR-6893-5p, hsa-miR-4728-5p, hsa-miR-6717-5p,hsa-miR-7113-3p, hsa-miR-4665-5p, hsa-miR-642b-3p, hsa-miR-7109-5p,hsa-miR-6842-5p, hsa-miR-4442, hsa-miR-4433-3p, hsa-miR-4707-5p,hsa-miR-6126, hsa-miR-4449, hsa-miR-4706, hsa-miR-1913, hsa-miR-602,hsa-miR-939-5p, hsa-miR-4695-5p, hsa-miR-711, hsa-miR-6816-5p,hsa-miR-4632-5p, hsa-miR-6721-5p, hsa-miR-7847-3p, hsa-miR-6132,hsa-miR-887-3p, hsa-miR-3679-3p, hsa-miR-6784-5p, hsa-miR-1249,hsa-miR-937-5p, hsa-miR-5195-3p, hsa-miR-6732-5p, hsa-miR-4417,hsa-miR-4281, hsa-miR-4734, hsa-miR-6766-3p, hsa-miR-663a, hsa-miR-4513,hsa-miR-6781-5p, hsa-miR-1227-5p, hsa-miR-6845-5p, hsa-miR-6798-5p,hsa-miR-3620-5p, hsa-miR-1915-5p, hsa-miR-4294, hsa-miR-642a-3p,hsa-miR-371a-5p, hsa-miR-940, hsa-miR-4450, hsa-miR-4723-5p,hsa-miR-1469, hsa-miR-6861-5p, hsa-miR-7975, hsa-miR-6879-5p,hsa-miR-6802-5p, hsa-miR-1268b, hsa-miR-663b, hsa-miR-125a-3p,hsa-miR-2861, hsa-miR-6088, hsa-miR-4758-5p, hsa-miR-296-3p,hsa-miR-6738-5p, hsa-miR-671-5p, hsa-miR-4454, hsa-miR-4516,hsa-miR-7845-5p, hsa-miR-4741, hsa-miR-92b-5p, hsa-miR-6795-5p,hsa-miR-6805-3p, hsa-miR-4725-3p, hsa-miR-6782-5p, hsa-miR-4688,hsa-miR-6850-5p, hsa-miR-6777-5p, hsa-miR-6785-5p, hsa-miR-7106-5p,hsa-miR-3663-3p, hsa-miR-6131, hsa-miR-1915-3p, hsa-miR-4532,hsa-miR-6820-5p, hsa-miR-4689, hsa-miR-4638-5p, hsa-miR-3656,hsa-miR-3621, hsa-miR-6769b-5p, hsa-miR-149-3p, hsa-miR-23b-3p,hsa-miR-3135b, hsa-miR-6848-5p, hsa-miR-6769a-5p, hsa-miR-4327,hsa-miR-6765-3p, hsa-miR-6716-5p, hsa-miR-6877-5p, hsa-miR-6727-5p,hsa-miR-4534, hsa-miR-614, hsa-miR-1202, hsa-miR-575, hsa-miR-6870-5p,hsa-miR-6722-3p, hsa-miR-7977, hsa-miR-4649-5p, hsa-miR-4675,hsa-miR-6075, hsa-miR-6779-5p, hsa-miR-4271, hsa-miR-3196,hsa-miR-6803-5p, hsa-miR-6789-5p, hsa-miR-4648, hsa-miR-4508,hsa-miR-4749-5p, hsa-miR-4505, hsa-miR-5698, hsa-miR-1199-5p andhsa-miR-4763-3p, hsa-miR-1231, hsa-miR-1233-5p, hsa-miR-150-3p,hsa-miR-1225-3p, hsa-miR-92a-2-5p, hsa-miR-423-5p, hsa-miR-1268a,hsa-miR-128-2-5p and hsa-miR-24-3p genes, and polynucleotides consistingof the nucleotide sequences of SEQ ID NOs: 1 to 180 related thereto werefound.

Among them, genes newly found as markers for examining the presence orabsence of colorectal cancer are polynucleotides consisting of thenucleotide sequences represented by SEQ ID NOs: 1 to 171.

A discriminant for determining the presence or absence of colorectalcancer was further prepared by Fisher's discriminant analysis with theexpression levels of these genes as an index. Specifically, anypolynucleotide consisting of a nucleotide sequence represented by any ofSEQ ID NOs: 1 to 180 found in the training cohort was applied to Formula2 above to construct a discriminant. Calculated accuracy, sensitivity,and specificity are shown in Table 3. In this respect, a discriminantcoefficient and a constant term are shown in Table 4.

Accuracy, sensitivity, and specificity in the validation cohort werecalculated using the discriminant thus prepared, and the discriminantperformance of the selected polynucleotides was validated usingindependent samples (Table 3). For example, the expression levelmeasurement value of the nucleotide sequence represented by SEQ ID NO: 1was compared between the healthy subjects (100 persons) and thecolorectal cancer patients (34 persons) in the training cohort. As aresult, the gene expression level measurement values were found to besignificantly lower in the colorectal cancer patient group than in thehealthy subject group (see the left diagram of FIG. 2 ). These resultswere also reproducible for the healthy subjects (50 persons) and thecolorectal cancer patients (16 persons) in the validation cohort (seethe right diagram of FIG. 2 ). Likewise, the results obtained about theother polynucleotides shown in SEQ ID NOs: 2 to 180 showed that the geneexpression level measurement values were significantly lower (−) orhigher (+) in the colorectal cancer patient group than in the healthysubject group (Table 2). These results were able to be validated in thevalidation cohort. For example, as for this nucleotide sequencerepresented by SEQ ID NO: 1, the number of samples that correctlyidentified in the detection of colorectal cancer in the validationcohort was calculated using the threshold (9.43) that was set in thetraining cohort and discriminated between the two groups. As a result,16 true positives, 50 true negatives, 0 false positives, and 0 falsenegatives were obtained. From these values, 100% accuracy, 100%sensitivity, and 100% specificity were obtained as detectionperformance. In this way, the detection performance was calculated as toall of the polynucleotides shown in SEQ ID NOs: 1 to 180, and describedin Table 3.

For example, 110 polynucleotides consisting of the nucleotide sequencesrepresented by SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13,14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31,32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 49, 50,51, 52, 53, 55, 56, 57, 58, 59, 60, 61, 62, 63, 65, 66, 67, 68, 69, 71,72, 73, 74, 76, 77, 78, 79, 80, 81, 83, 84, 86, 87, 88, 90, 92, 93, 95,96, 97, 99, 100, 101, 102, 107, 109, 110, 111, 113, 114, 115, 118, 120,122, 124, 126, 134, 136, 142, 153, 172, 173 and 175 exhibitedsensitivity of 100%, 100%, 100%, 75%, 93.8%, 75%, 87.5%, 75%, 93.8%,68.8%, 81.2%, 100%, 75%, 50%, 75%, 75%, 68.8%, 75%, 81.2%, 81.2%, 75%,62.5%, 75%, 56.2%, 75%, 68.8%, 56.2%, 62.5%, 68.8%, 75%, 68.8%, 68.8%,56.2%, 68.8%, 62.5%, 68.8%, 62.5%, 50%, 56.2%, 56.2%, 56.2%, 75%, 50%,68.8%, 68.8%, 68.8%, 50%, 56.2%, 62.5%, 62.5%, 50%, 62.5%, 68.8%, 56.2%,56.2%, 43.8%, 75%, 62.5%, 62.5%, 56.2%, 62.5%, 62.5%, 56.2%, 62.5%,56.2%, 56.2%, 56.2%, 56.2%, 43.8%, 43.8%, 50%, 68.8%, 56.2%, 62.5%,62.5%, 43.8%, 62.5%, 56.2%, 62.5%, 62.5%, 50%, 56.2%, 43.8%, 50%, 43.8%,50%, 43.8%, 56.2%, 43.8%, 50%, 50%, 50%, 50%, 50%, 50%, 43.8%, 50%,43.8%, 50%, 50%, 50%, 43.8%, 43.8%, 50%, 43.8%, 43.8%, 50%, 81.2%, 68.8%and 56.2%, respectively in the validation cohort (Table 3). As seen fromComparative Example mentioned later, the existing markers CEA hadsensitivity of 43.75% in the validation cohort (Tables 5-1 and 5-2),demonstrating that the 110 polynucleotides consisting of the nucleotidesequences represented by SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11,12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29,30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47,49, 50, 51, 52, 53, 55, 56, 57, 58, 59, 60, 61, 62, 63, 65, 66, 67, 68,69, 71, 72, 73, 74, 76, 77, 78, 79, 80, 81, 83, 84, 86, 87, 88, 90, 92,93, 95, 96, 97, 99, 100, 101, 102, 107, 109, 110, 111, 113, 114, 115,118, 120, 122, 124, 126, 134, 136, 142, 153, 172, 173 and 175 candiscriminate, each alone, colorectal cancer in the validation cohortwith sensitivity beyond CEA.

For example, 14 polynucleotides consisting of the nucleotide sequencesrepresented by SEQ ID NOs: 1, 2, 3, 10, 14, 17, 21, 23, 32, 36, 47, 59,65, and 101 were able to correctly determine colorectal cancer as to allof three stage 1 colorectal cancer samples that were contained in thevalidation cohort. Thus, these polynucleotides can detect even earlycolorectal cancer and contribute to the early diagnosis of colorectalcancer.

For example, 12 polynucleotides consisting of the nucleotide sequencesrepresented by SEQ ID NOs: 1, 2, 3, 5, 7, 10, 14, 39, 46, 73, 81, and148 were able to correctly determine colorectal cancer as to all of onececal cancer case and 3 ascending colon cancer cases, which were cancercases in the upper large intestine that are reportedly difficult todetect by the fecal occult blood test, in the validation cohort. Thus,these polynucleotides can detect colorectal cancer regardless of wherecolorectal cancer develops.

Example 2

<Method for Evaluating Colorectal Cancer Discriminant Performance byCombination of Multiple Gene Markers Using Samples in the ValidationCohort>

In this Example, a method for evaluating colorectal cancer discriminantperformance by a combination of the gene markers selected in Example 1was studied. Specifically, Fisher's discriminant analysis was conductedas to 16,074 combinations of two polynucleotides comprising at least oneor more of the expression level measurement values of the newly foundpolynucleotides consisting of the nucleotide sequences represented bySEQ ID NOs: 1 to 171 among the polynucleotides consisting of thenucleotide sequences represented by SEQ ID NOs: 1 to 180 selected inExample 1, to construct a discriminant for determining the presence orabsence of colorectal cancer. Next, accuracy, sensitivity, andspecificity in the validation cohort were calculated using thediscriminant thus prepared, and the discriminant performance of theselected polynucleotides was validated using independent samples.

For example, the expression level measurement values of the nucleotidesequences represented by SEQ ID NO: 1 and SEQ ID NO: 2 were comparedbetween the healthy subjects (100 persons) and the colorectal cancerpatients (34 persons) in the training cohort. As a result, a scatterdiagram that significantly separated the gene expression levelmeasurement values of the colorectal cancer patient group from those ofthe healthy subject group was obtained (see the left diagram of FIG. 3). These results were also reproducible for the healthy subjects (50persons) and the colorectal cancer patients (16 persons) in thevalidation cohort (see the right diagram of FIG. 3 ). Likewise, ascatter diagram that significantly separated the gene expression levelmeasurement values of the colorectal cancer patient group from those ofthe healthy subject group was also obtained as to the other combinationsof two expression level measurement values comprising at least one ormore of the expression level measurement values of the newly foundpolynucleotides consisting of the nucleotide sequences represented bySEQ ID NOs: 1 to 171 among the polynucleotides consisting of thenucleotide sequences represented by SEQ ID NOs: 1 to 180. These resultswere able to be validated in the validation cohort. For example, as forthese nucleotide sequences represented by SEQ ID NO: 1 and SEQ ID NO: 2,the number of samples that correctly identified in the detection ofcolorectal cancer was calculated using the function (0=1.26x+y−18.06)that was set in the training cohort and discriminated between the twogroups. As a result, 16 true positives, 50 true negatives, 0 falsepositives, and 0 false negatives were obtained. From these values, 100%accuracy, 100% sensitivity, and 100% specificity were obtained asdetection performance. In this way, the detection performance wascalculated as to all of the combinations of two expression levelmeasurement values comprising at least one or more of the expressionlevel measurement values of the newly found polynucleotides consistingof the nucleotide sequences represented by SEQ ID NOs: 1 to 171 amongthe polynucleotides consisting of the nucleotide sequences representedby SEQ ID NOs: 1 to 180. Among them, 179 combinations comprising theexpression level measurement value of the polynucleotide consisting ofthe nucleotide sequence represented by SEQ ID NO: 1 and the detectionperformance thereof were described in Table 6 as an example. Forexample, all of combinations of the expression level measurement valuesof the polynucleotides consisting of the nucleotide sequencesrepresented by SEQ ID NOs: 1 and 2, SEQ ID NOs: 1 and 3, SEQ ID NOs: 1and 4, and SEQ ID NOs: 1 and 5 exhibited sensitivity of 100% in thevalidation cohort (Table 6). Further, combinations of twopolynucleotides consisting of nucleotide sequences other than SEQ ID NO:1 were described in Table 7 as an example. As specific combinations oftwo polynucleotides, for example, combinations represented by SEQ IDNOs: 5 and 6, SEQ ID NOs: 5 and 11, SEQ ID NOs: 5 and 38, SEQ ID NOs: 15and 16, SEQ ID NOs: 15 and 21, SEQ ID NOs: 15 and 64, SEQ ID NOs: 24 and25, SEQ ID NOs: 24 and 30, SEQ ID NOs: 24 and 32, SEQ ID NOs: 2 and 32,SEQ ID NOs: 32 and 36, SEQ ID NOs: 15 and 32, SEQ ID NOs: 3 and 38, SEQID NOs: 38 and 39, SEQ ID NOs: 38 and 64, SEQ ID NOs: 3 and 45, SEQ IDNOs: 45 and 58, SEQ ID NOs: 45 and 64, SEQ ID NOs: 2 and 55, SEQ ID NOs:6 and 55, SEQ ID NOs: 55 and 64, SEQ ID NOs: 2 and 64, SEQ ID NOs: 4 and64, SEQ ID NOs: 2 and 96, SEQ ID NOs: 7 and 96, SEQ ID NOs: 96 and 97,SEQ ID NOs: 2 and 97, SEQ ID NOs: 3 and 97, SEQ ID NOs: 5 and 97, SEQ IDNOs: 2 and 162, SEQ ID NOs: 3 and 162, and SEQ ID NOs: 5 and 162,exhibited accuracy of 75% or higher for discriminating the colorectalcancer patients from the healthy subjects in both of the training cohortand the validation cohort. In this way, 14,598 combinations of theexpression level measurement values of two polynucleotides that havesensitivity beyond the existing marker CEA (43.8% in Table 5-2) wereobtained in the validation cohort. All of the nucleotide sequences 1 to180 described in Table 2 obtained in Example 1 were employed at leastonce in these combinations. These results demonstrated that the combineduse of two of the polynucleotides consisting of the nucleotide sequencesrepresented by SEQ ID NOs: 1 to 180 can also discriminate colorectalcancer with excellent performance beyond the existing marker. Markersfor the detection of colorectal cancer with better sensitivity areobtained by combining 3, 4, 5, 6, 7, 8, 9, 10 or more of the expressionlevel measurement values of the polynucleotides consisting of thenucleotide sequences represented by SEQ ID NOs: 1 to 180. For example,the newly found polynucleotides consisting of the nucleotide sequencesrepresented by SEQ ID NOs: 1 to 171 among the polynucleotides consistingof the nucleotide sequences represented by SEQ ID NOs: 1 to 180 selectedin Example 1 were measured to obtain their expression levels between thehealthy subject group and the colorectal cancer group in the validationcohort. All of the polynucleotides were ranked in the descending orderof their P values obtained by the Student's t-test, which indicatesstatistical significance of difference between groups (i.e., one havingthe lowest P value was ranked in the first place), and colorectal cancerdetection sensitivity was evaluated using combinations of one or morepolynucleotides to which the polynucleotides were added to thecombination one by one from the top to the bottom according to the rank.In short, the order in which the polynucleotides were combined in thisevaluation is in reverse in terms of SEQ ID NOs, from SEQ ID NO: 171 toSEQ ID NOs: 170, 169, . . . as shown in Table 2. As a result, thesensitivity in the validation cohort was 12.5% for 1 polynucleotide (SEQID NO: 171), 18.8% for 2 polynucleotides (SEQ ID NOs: 170 and 171),25.0% for 4 polynucleotides (SEQ ID NOs: 168 to 171), 31.2% for 5polynucleotides (SEQ ID NOs: 167 to 171), 37.5% for 7 polynucleotides(SEQ ID NOs: 165 to 171), 87.5% for 10 polynucleotides (SEQ ID NOs: 162to 171), 100% for 20 polynucleotides (SEQ ID NOs: 152 to 171), 100% for30 polynucleotides (SEQ ID NOs: 142 to 171), 100% for 80 polynucleotides(SEQ ID NOs: 92 to 171), 100% for 170 polynucleotides (SEQ ID NOs: 2 to171), and 100% for 171 polynucleotides (SEQ ID NOs: 1 to 171).

These results demonstrated that a combination of multiplepolynucleotides can produce higher colorectal cancer discriminantperformance than that of each polynucleotide alone or a combination of afewer number of polynucleotides. In this context, the combinations ofmultiple polynucleotides are not limited to the combinations of thepolynucleotides added in the order of statistically significantdifference as described above, and any combination of multiplepolynucleotides can be used in the detection of colorectal cancer.

From these results, it can be concluded that all of the polynucleotidesconsisting of the nucleotide sequences represented by SEQ ID NOs: 1 to180 serve as excellent markers for the detection of colorectal cancer.

TABLE 2 Expression level in P value after colorectal cancer Bonferronipatient with respect SEQ ID NO: Gene name correction to healthy subject1 hsa-miR-6726-5p 5.20.E−41 − 2 hsa-miR-4257 7.54.E−40 − 3hsa-miR-6787-5p 1.72.E−30 − 4 hsa-miR-6780b-5p 3.42.E−30 + 5hsa-miR-3131 1.62.E−27 − 6 hsa-miR-7108-5p 5.42.E−27 + 7 hsa-miR-1343-3p2.12.E−26 − 8 hsa-miR-1247-3p 9.98.E−26 + 9 hsa-miR-4651 3.90.E−24 − 10hsa-miR-6757-5p 2.25.E−23 − 11 hsa-miR-3679-5p 2.55.E−23 + 12hsa-miR-7641 971.E−22 − 13 hsa-miR-6746-5p 1.64.E−21 − 14 hsa-miR-80724.09.E−21 + 15 hsa-miR-6741-5p 7.23.E−21 − 16 hsa-miR-1908-5p2.12.E−20 + 17 hsa-miR-6857-5p 2.70.E−20 + 18 hsa-miR-4746-3p3.58.E−20 + 19 hsa-miR-744-5p 4.23.E−20 + 20 hsa-miR-4792 8.25.E−20 + 21hsa-miR-564 1.78.E−19 − 22 hsa-miR-6791-5p 3.80.E−19 + 23hsa-miR-6825-5p 5.93.E−19 + 24 hsa-miR-6826-5p 8.67.E−19 − 25hsa-miR-4665-3p 1.92.E−18 + 26 hsa-miR-4467 5.55.E−18 + 27 hsa-miR-31888.48.E−18 + 28 hsa-miR-6125 1.09.E−17 + 29 hsa-miR-6756-5p 1.24.E−17 −30 hsa-miR-1228-3p 1.68.E−17 + 31 hsa-miR-8063 2.70.E−17 − 32hsa-miR-8069 3.58.E−17 + 33 hsa-miR-6875-5p 6.07.E−17 + 34 hsa-miR-31855.07.E−16 + 35 hsa-miR-4433b-3p 1.22.E−15 + 36 hsa-miR-6887-5p 1.30.E−15− 37 hsa-miR-128-1-5p 3.61.E−15 + 38 hsa-miR-6724-5p 3.81.E−15 + 39hsa-miR-1914-3p 1.05.E−14 − 40 hsa-miR-1225-5p 3.93.E−14 + 41hsa-miR-4419b 5.90.E−14 − 42 hsa-miR-7110-5p 6.01.E−14 + 43hsa-miR-187-5p 8.57.E−14 − 44 hsa-miR-3184-5p 1.40.E−13 + 45hsa-miR-204-3p 2.23.E−13 − 46 hsa-miR-5572 2.34.E−13 + 47hsa-miR-6729-5p 3.33.E−13 + 48 hsa-miR-615-5p 4.27.E−13 − 49hsa-miR-6749-5p 5.30.E−13 − 50 hsa-miR-6515-3p 7.31.E−13 + 51hsa-miR-3937 8.10.E−13 + 52 hsa-miR-6840-3p 1.15.E−12 − 53hsa-miR-6893-5p 1.34.E−12 − 54 hsa-miR-4728-5p 2.48.E−12 − 55hsa-miR-6717-5p 4.45.E−12 − 56 hsa-miR-7113-3p 5.11.E−12 + 57hsa-miR-4665-5p 5.33.E−12 − 58 hsa-miR-642b-3p 6.74.E−12 − 59hsa-miR-7109-5p 6.88.E−12 − 60 hsa-miR-6842-5p 6.91.E−12 + 61hsa-miR-4442 8.87.E−12 − 62 hsa-miR-4433-3p 9.88.E−12 + 63hsa-miR-4707-5p 1.19.E−11 + 64 hsa-miR-6126 1.27.E−11 + 65 hsa-miR-44491.32.E−11 + 66 hsa-miR-4706 2.85.E−11 − 67 hsa-miR-1913 3.15.E−11 + 68hsa-miR-602 4.98.E−11 + 69 hsa-miR-939-5p 6.08.E−11 + 70 hsa-miR-4695-5p8.15.E−11 + 71 hsa-miR-711 1.23 E−10 + 72 hsa-miR-6816-5p 1.29.E−10 + 73hsa-miR-4632-5p 1.50.E−10 + 74 hsa-miR-6721-5p 1.98.E−10 + 75hsa-miR-7847-3p 2.14.E−10 − 76 hsa-miR-6132 2.68.E−10 + 77hsa-miR-887-3p 2.81.E−10 + 78 hsa-miR-3679-3p 3.07.E−10 + 79hsa-miR-6784-5p 3.20.E−10 + 80 hsa-miR-1249 3.40.E−10 + 81hsa-miR-937-5p 5.57.E−10 − 82 hsa-miR-5195-3p 6.88.E−10 − 83hsa-miR-6732-5p 7.27.E−10 + 84 hsa-miR-4417 7.95.E−10 + 85 hsa-miR-42819.35.E−10 − 86 hsa-miR-4734 1.04.E−09 + 87 hsa-miR-6766-3p 1.07.E−09 +88 hsa-miR-663a 2.19.E−09 + 89 hsa-miR-4513 3.03.E−09 − 90hsa-miR-6781-5p 5.11.E−09 + 91 hsa-miR-1227-5p 6.16.E−09 + 92hsa-miR-6845-5p 6.49.E−09 + 93 hsa-miR-6798-5p 8.99.E−09 + 94hsa-miR-3620-5p 1.09.E−08 + 95 hsa-miR-1915-5p 1.78.E−08 − 96hsa-miR-4294 2.30.E−08 − 97 hsa-miR-642a-3p 2.61.E−08 − 98hsa-miR-371a-5p 3.15.E−08 − 99 hsa-miR-940 3.18.E−08 + 100 hsa-miR-44503.25.E−08 − 101 hsa-miR-4723-5p 4.21.E−08 − 102 hsa-miR-1469 4.26.E−08 +103 hsa-miR-6861-5p 4.71.E−08 − 104 hsa-miR-7975 7.28.E−08 − 105hsa-miR-6879-5p 7.64.E−08 + 106 hsa-miR-6802-5p 9.22.E−08 − 107hsa-miR-1268b 1.08.E−07 + 108 hsa-miR-663b 1.12.E−07 − 109hsa-miR-125a-3p 1.16.E−07 − 110 hsa-miR-2861 1.87.E−07 − 111hsa-miR-6088 2.97.E−07 − 112 hsa-miR-4758-5p 3.12.E−07 − 113hsa-miR-296-3p 3.43.E−07 − 114 hsa-miR-6738-5p 4.05.E−07 − 115hsa-miR-671-5p 5.76.E−07 − 116 hsa-miR-4454 6.68.E−07 − 117 hsa-miR-45161.04.E−06 − 118 hsa-miR-7845-5p 1.10.E−06 + 119 hsa-miR-4741 1.52.E−06 +120 hsa-miR-92b-5p 1.63.E−06 + 121 hsa-miR-6795-5p 2.31.E−06 − 122hsa-miR-6805-3p 3.95.E−06 + 123 hsa-miR-4725-3p 5.35.E−06 + 124hsa-miR-6782-5p 5.69.E−06 + 125 hsa-miR-4688 8.95.E−06 − 126hsa-miR-6850-5p 1.66.E−05 + 127 hsa-miR-6777-5p 1.74.E−05 − 128hsa-miR-6785-5p 1.89.E−05 − 129 hsa-miR-7106-5p 1.94.E−05 − 130hsa-miR-3663-3p 2.08.E−05 − 131 hsa-miR-6131 2.29.E−05 − 132hsa-miR-1915-3p 3.16.E−05 + 133 hsa-miR-4532 3.46.E−05 − 134hsa-miR-6820-5p 3.81.E−05 − 135 hsa-miR-4689 4.54.E−05 − 136hsa-miR-4638-5p 4.70.E−05 − 137 hsa-miR-3656 5.75.E−05 + 138hsa-miR-3621 6.34.E−05 − 139 hsa-miR-6769b-5p 6.63.E−05 − 140hsa-miR-149-3p 1.01.E−04 − 141 hsa-miR-23b-3p 1.11.E−04 − 142hsa-miR-3135b 1.16.E−04 − 143 hsa-miR-6848-5p 1.17.E−04 + 144hsa-miR-6769a-5p 1.23.E−04 − 145 hsa-miR-4327 1.40.E−04 + 146hsa-miR-6765-3p 1.50.E−04 − 147 hsa-miR-6716-5p 1.51.E−04 + 148hsa-miR-6877-5p 1.52.E−04 − 149 hsa-miR-6727-5p 2.04.E−04 − 150hsa-miR-4534 2.10.E−04 − 151 hsa-miR-614 3.18.E−04 − 152 hsa-miR-12024.86.E−04 − 153 hsa-miR-575 4.92.E−04 − 154 hsa-miR-6870-5p 5.55.E−04 +155 hsa-miR-6722-3p 7.07.E−04 + 156 hsa-miR-7977 7.17.E−04 − 157hsa-miR-4649-5p 7.70.E−04 − 158 hsa-miR-4675 9.21.E−04 − 159hsa-miR-6075 1.03.E−03 + 160 hsa-miR-6779-5p 1.04.E−03 − 161hsa-miR-4271 1.43.E−03 − 162 hsa-miR-3196 1.45.E−03 + 163hsa-miR-6803-5p 1.46.E−03 + 164 hsa-miR-6789-5p 1.71.E−03 + 165hsa-miR-4648 1.90.E−03 + 166 hsa-miR-4508 3.41.E−03 + 167hsa-miR-4749-5p 3.52.E−03 + 168 hsa-miR-4505 4.01.E−03 + 169hsa-miR-5698 4.99.E−03 − 170 hsa-miR-1199-5p 5.88.E−03 − 171hsa-miR-4763-3p 8.40.E−03 + 172 hsa-miR-1231 7.36.E−25 + 173hsa-miR-1233-5p 1.21.E−22 − 174 hsa-miR-150-3p 5.76.E−07 − 175hsa-miR-1225-3p 1.44.E−06 + 176 hsa-miR-92a-2-5p 2.36.E−05 + 177hsa-miR-423-5p 4.62.E−05 − 178 hsa-miR-1268a 4.30.E−04 + 179hsa-miR-128-2-5p 6.64.E−04 − 180 hsa-miR-24-3p 1.31.E−03 −

TABLE 3 Training cohort Validation cohort Accuracy SensitivitySpecificity Accuracy Sensitivity Specificity SEQ ID NO: (9) (%) (%) (%)(%) (%) 1 99.3 100 99 100 100 100 2 96.3 88.2 99 100 100 100 3 96.3 91.298 98.5 100 98 4 93.3 85.3 96 93.9 75 100 5 97 91.2 99 97 93.8 98 6 9482.4 98 90.9 75 96 7 96.3 88.2 99 95.5 87.5 98 8 92.5 82.4 96 89.4 75 949 93.3 85.3 96 97 93.8 98 10 91.8 79.4 96 92.4 68.8 100 11 94.8 91.2 9695.5 81.2 100 12 90.3 82.4 93 97 100 96 13 89.6 79.4 93 90.9 75 96 14 9173.5 97 80.3 50 90 15 94 79.4 99 89.4 75 94 16 88.1 73.5 93 89.4 75 9417 91 85.3 93 87.9 68.8 94 18 91 79.4 95 92.4 75 98 19 90.3 76.5 95 93.981.2 98 20 91.8 88.2 93 92.4 81.2 96 21 87.3 58.8 97 92.4 75 98 22 88.173.5 93 89.4 62.5 98 23 87.3 79.4 90 87.9 75 92 24 90.3 67.6 98 89.456.2 100 25 89.6 67.6 97 84.8 75 88 26 83.6 70.6 88 89.4 68.8 96 27 91.876.5 97 87.9 56.2 98 28 91 82.4 94 87.9 62.5 96 29 88.8 67.6 96 83.368.8 88 30 91.8 85.3 94 86.4 75 90 31 87.3 79.4 90 87.9 68.8 94 32 87.364.7 95 89.4 68.8 96 33 91 79.4 95 80.3 56.2 88 34 89.6 76.5 94 89.468.8 96 35 89.6 79.4 93 78.8 62.5 84 36 88.1 55.9 99 92.4 68.8 100 3785.1 61.8 93 80.3 62.5 86 38 86.6 70.6 92 78.8 50 88 39 88.1 70.6 9481.8 56.2 90 40 91 76.5 96 84.8 56.2 94 41 86.6 58.8 96 87.9 56.2 98 4284.3 64.7 91 86.4 75 90 43 84.3 52.9 95 86.4 50 98 44 87.3 70.6 93 87.968.8 94 45 87.3 61.8 96 77.3 68.8 80 46 83.6 70.6 88 84.8 68.8 90 4786.6 52.9 98 86.4 50 98 48 88.8 58.8 99 81.8 31.2 98 49 87.3 61.8 9687.9 56.2 98 50 86.6 73.5 91 77.3 62.5 82 51 86.6 64.7 94 87.9 62.5 9652 84.3 52.9 95 84.8 50 96 53 88.8 64.7 97 87.9 62.5 96 54 81.3 50 9277.3 31.2 92 55 88.8 58.8 99 90.9 68.8 98 56 84.2 66.7 90 83.3 56.2 9257 84.3 58.8 93 80.3 56.2 88 58 85.1 50 97 86.4 43.8 100 59 82.8 55.9 9289.4 75 94 60 87.3 64.7 95 87.9 62.5 96 61 81.3 52.9 91 84.8 62.5 92 6282.8 67.6 88 80.3 56.2 88 63 82.1 55.9 91 84.8 62.5 92 64 78.4 38.2 9283.3 37.5 98 65 86.6 61.8 95 87.9 62.5 96 66 85.1 58.8 94 84.8 56.2 9467 83.6 61.8 91 80 62.5 85.7 68 85.1 61.8 93 84.8 56.2 94 69 80.6 64.786 80.3 56.2 88 70 81.3 52.9 91 78.8 31.2 94 71 85.1 58.8 94 87.9 56.298 72 83.6 64.7 90 83.3 56.2 92 73 87.3 55.9 98 84.8 43.8 98 74 83.664.7 90 77.3 43.8 88 75 82.7 33.3 99 84.8 37.5 100 76 83.6 44.1 97 86.450 98 77 85.8 73.5 90 83.3 68.8 88 78 83.6 52.9 94 81.8 56.2 90 79 83.667.6 89 81.8 62.5 88 80 85 58.8 93.9 83.3 62.5 90 81 84.3 50 96 83.343.8 96 82 81.3 44.1 94 81.8 37.5 96 83 82.1 61.8 89 78.8 62.5 84 8490.3 70.6 97 84.8 56.2 94 85 83.6 55.9 93 80.3 31.2 96 86 80.6 41.2 9486.4 62.5 94 87 83.6 50 95 83.3 62.5 90 88 84.3 52.9 95 83.3 50 94 8984.3 44.1 98 77.3 12.5 98 90 82.8 50 94 81.8 56.2 90 91 79.9 38.2 9475.8 31.2 90 92 84.3 50 96 78.8 43.8 90 93 82.8 61.8 90 75.8 50 84 9484.3 55.9 94 77.3 31.2 92 95 82.1 41.2 96 83.3 43.8 96 96 85.1 55.9 9581.8 50 92 97 78.4 38.2 92 78.8 43.8 90 98 82.8 50 94 75.8 37.5 88 9981.3 47.1 93 86.4 56.2 96 100 85.1 47.1 98 83.3 43.8 96 101 87.3 58.8 9783.3 50 94 102 80.6 38.2 95 80.3 50 90 103 83.6 47.1 96 80.3 37.5 94 10479.1 35.3 94 78.8 37.5 92 105 82.8 38.2 98 84.8 37.5 100 106 82.8 44.196 81.8 37.5 96 107 74.6 32.4 89 75.8 50 84 108 83.6 47.1 96 83.3 31.2100 109 85.1 44.1 99 87.9 50 100 110 82.8 52.9 93 84.8 50 96 111 78.444.1 90 81.8 50 92 112 84.3 44.1 98 80.3 25 98 113 82.8 50 94 80.3 43.892 114 82.8 52.9 93 83.3 50 94 115 82.1 44.1 95 84.8 43.8 98 116 79.941.2 93 77.3 31.2 92 117 87.3 50 100 84.8 37.5 100 118 88.1 58.8 98 81.850 92 119 78.4 29.4 95 77.3 25 94 120 78.4 41.2 91 84.8 50 96 121 80.626.5 99 80.3 18.8 100 122 77.6 38.2 91 83.3 50 94 123 76.1 26.5 93 74.212.5 94 124 83.6 44.1 97 83.3 43.8 96 125 77.6 35.3 92 74.2 18.8 92 12680.6 41.2 94 78.8 43.8 90 127 79.1 23.5 98 83.3 31.2 100 128 80.6 38.295 80.3 31.2 96 129 78.4 23.5 97 80.3 25 98 130 78.4 29.4 95 80.3 31.296 131 81.3 35.3 97 83.3 37.5 98 132 80.6 35.3 96 80.3 25 98 133 82.844.1 96 80.3 37.5 94 134 83.6 41.2 98 83.3 50 94 135 79.9 29.4 97 81.825 100 136 83.6 41.2 98 86.4 43.8 100 137 79.9 38.2 94 77.3 12.5 98 13876.1 26.5 93 77.3 25 94 139 79.1 26.5 97 78.8 18.8 98 140 76.9 23.5 9577.3 25 94 141 79.1 26.5 97 75.8 18.8 94 142 83.6 38.2 99 86.4 43.8 100143 77.6 26.5 95 78.8 25 96 144 74.6 17.6 94 80.3 31.2 96 145 79.1 41.292 75.8 25 92 146 78.4 32.4 94 80.3 31.2 96 147 79.1 29.4 96 77.3 31.292 148 73.9 20.6 92 71.2 6.2 92 149 79.1 38.2 93 81.8 31.2 98 150 78.423.5 97 74.2 25 90 151 76.1 32.4 91 77.3 25 94 152 81.3 29.4 99 81.8 25100 153 82.1 29.4 100 87.9 50 100 154 81.3 35.3 97 84.8 37.5 100 15579.1 29.4 96 78.8 31.2 94 156 78.9 24.2 97 77.3 25 94 157 79.9 29.4 9783.3 31.2 100 158 80.6 35.3 96 84.8 37.5 100 159 82.1 35.3 98 81.8 31.298 160 78.4 20.6 98 81.8 31.2 98 161 78.4 26.5 96 81.8 25 100 162 79.129.4 96 77.3 18.8 96 163 74.6 26.5 91 63.6 0 84 164 76.1 20.6 95 71.212.5 90 165 77.6 23.5 96 81.8 25 100 166 78.4 29.4 95 69.7 6.2 90 16778.4 14.7 100 75.8 0 100 168 78.2 21.2 97 78.8 12.5 100 169 78.4 23.5 9777.3 6.2 100 170 73.9 2.9 98 77.3 6.2 100 171 80.6 26.5 99 78.8 12.5 100172 93.3 85.3 96 90.9 81.2 94 173 91 76.5 96 90.9 68.8 98 174 82.1 35.398 77.3 31.2 92 175 87.3 52.9 99 89.4 56.2 100 176 74.6 29.4 90 78.837.5 92 177 79.9 35.3 95 69.7 12.5 88 178 73.9 17.6 93 71.2 6.2 92 17981.3 32.4 98 84.8 37.5 100 180 76.9 11.8 99 81.8 25 100

TABLE 4 Discriminant Constant SEQ ID NO: coefficient term 1 3.451 32.5372 2.778 17.111 3 3.893 32.032 4 3.208 29.340 5 2.408 15.716 6 4.76044.132 7 1.872 13.040 8 4.189 26.554 9 5.692 61.192 10 2.915 20.140 112.801 19.585 12 1.247 8.323 13 3.434 21.316 14 5.315 65.956 15 3.97126.352 16 4.335 50.272 17 1.843 9.956 18 2.796 18.550 19 2.726 19.273 202.151 14.586 21 1.432 7.567 22 4.810 44.500 23 2.202 14.554 24 1.7879.999 25 4.048 23.773 26 2.353 23.473 27 3.139 19.203 28 5.364 64.417 295.274 42.891 30 4.406 27.813 31 2.590 20.814 32 6.586 84.911 33 3.42631.099 34 2.365 16.821 35 3.810 30.817 36 2.245 13.547 37 2.667 20.06038 4.817 48.162 39 4.582 33.609 40 3.409 25.092 41 2.180 12.620 42 1.84614.493 43 2.092 20.352 44 2.237 18.151 45 1.808 22.979 46 2.361 15.74747 8.658 108.735 48 1.910 11.860 49 4.384 43.382 50 4.476 30.075 514.069 35.285 52 2.888 24.905 53 2.016 16.544 54 4.690 32.139 55 2.20713.044 56 3.152 18.319 57 3.384 31.679 58 2.167 19.956 59 5.078 36.90760 3.628 21.525 61 3.373 31.520 62 3.836 28.118 63 4.332 31.744 64 2.94932.215 65 3.709 24.031 66 3.738 28.272 67 3.638 22.448 68 3.013 19.23269 2.461 18.582 70 4.311 32.255 71 3.548 29.298 72 4.499 45.352 73 4.07932.445 74 3.995 30.128 75 2.483 15.148 76 3.479 27.463 77 2.342 16.97578 3.352 20.098 79 3.684 46.309 80 3.835 22.808 81 3.983 32.779 82 2.90419.401 83 3.426 29.138 84 5.296 43.216 85 3.793 43.429 86 5.582 66.47887 3.815 22.562 88 4.509 45.905 89 2.269 12.804 90 5.547 57.838 91 6.32560.270 92 3.946 37.787 93 2.967 30.962 94 3.865 30.606 95 1.266 7.550 962.410 24.206 97 2.733 20.281 98 3.561 25.772 99 3.064 19.551 100 1.1886.373 101 2.565 22.283 102 5.084 51.748 103 3.700 26.315 104 2.22421.832 105 3.135 25.894 106 4.526 37.574 107 3.166 31.384 108 2.83924.460 109 1.007 6.029 110 5.545 68.155 111 3.299 33.145 112 6.27153.263 113 2.148 12.402 114 3.608 25.322 115 2.758 17.059 116 2.17525.025 117 3.823 49.903 118 2.725 18.024 119 3.890 38.378 120 3.50627.825 121 2.582 15.075 122 2.476 18.382 123 4.084 39.823 124 2.97818.190 125 3.980 27.914 126 5.916 67.040 127 2.075 13.104 128 2.31720.667 129 2.093 12.035 130 4.219 50.899 131 1.841 19.246 132 3.96043.646 133 3.277 38.660 134 2.733 19.515 135 3.239 30.244 136 1.4828.655 137 4.554 52.325 138 5.175 61.317 139 3.430 21.115 140 5.43050.527 141 1.168 6.718 142 2.311 17.824 143 4.599 33.779 144 3.92124.668 145 4.968 43.118 146 1.700 14.753 147 3.593 23.332 148 4.30730.486 149 6.087 77.329 150 2.704 17.759 151 1.757 11.661 152 2.63516.886 153 1.214 6.968 154 3.201 23.463 155 6.593 55.857 156 2.17721.212 157 2.411 24.700 158 2.636 19.709 159 3.045 25.772 160 5.59339.283 161 3.606 29.381 162 6.360 76.890 163 6.727 74.567 164 4.35042.883 165 1.256 7.389 166 6.503 84.138 167 3.665 29.142 168 4.23335.592 169 1.766 10.169 170 1.955 12.693 171 3.328 27.665 172 3.67424.498 173 2.869 31.161 174 1.758 11.388 175 2.132 11.850 176 2.14820.104 177 2.169 15.443 178 3.124 34.907 179 2.552 27.422 180 1.4178.536

TABLE 5-1 Training cohort Sample name Cancer stage CEA(ng/mL)CA19-9(U/mL) CC03 I 1.6  13.5  CC04 I 2    30.6  CC05 I 1.3   3.2 CC06 I1.7  13.5  CC07 IIIA 4.4   0.1 CC09 IIIB 0.9   4.4 CC10 I 1.5  13.2 CC12 I 0.9  13.2  CC13 I 0.8   3.1 CC15 I 1.6   5.6 CC17 IIIA 2.7  21.7 CC18 I 3.2  16.4  CC19 IVL 6.2  45.9  CC20 IIIC 9.4   5.4 CC23 I 2.3  7.9 CC24 IIA 8.8  106.7   CC25 IIA 6.2  29.6  CC26 I 4.5  18.6  CC27IIIC 17.3    14.4  CC29 IIA 2.1   6.9 CC30 IIIA 3.2  13.2  CC31 IIIB6     5.7 CC32 IIIA 2.4  26.7  CC34 I 0.6   9.3 CC36 I 6.7   0.1 CC38IIA 1.2   6.1 CC40 IIIB 2.1   7.6 CC41 I 2.8  10.6  CC42 IIIB 46.7   3524   CC45 I 2.2  38.4  CC47 IIIB 1.7   7.1 CC48 IIA 2    19.1  CC49IIIB 0.9   8.1 CC50 IIA 7.6  12.2  Sensitivity 26.5%    12%  

TABLE 5-2 Validation cohort Sample name Cancer stage CEA(ng/mL)CA19-9(U/mL) CC0l I 2.2 13.9 CC02 I 3.9 16 CC08 IVH 15.4 9.5 CC11 IIIC7.2 8 CC14 I 0.6 14 CC16 IVL 10.1 106.7 CC21 IIIB 6.7 23.6 CC22 IIIC 2.942.4 CC28 IIIB 35.5 71 CC33 IIB 5 — CC35 IVH 20.3 552 CC37 IIA 0.1 8.1CC39 IVHLu 267.7 269.6 CC43 IIA 2 10.3 CC44 IIA 3.7 14 CC46 IIA 1.7 4.2Sensitivity   43.8%    31%

TABLE 6 Training cohort Validation cohort Accuracy SensitivitySpecificity Accuracy Sensitivity Specificity SEQ ID NO: (%) (%) (%) (%)(%) (%) 1_2 100 100 100 100 100 100 1_3 99.3 97.1 100 100 100 100 1_4100 100 100 100 100 100 1_5 100 100 100 100 100 100 1_6 100 100 100 100100 100 1_7 100 100 100 100 100 100 1_8 100 100 100 100 100 100 1_9 98.5100 98 98.5 100 98 1_10 100 100 100 100 100 100 1_11 99.3 100 99 100 100100 1_12 99.3 100 99 98.5 100 98 1_13 99.3 100 99 100 100 100 1_14 100100 100 98.5 100 98 1_15 99.3 97.1 100 97 100 96 1_16 100 100 100 100100 100 1_17 97.8 94.1 99 100 100 100 1_18 99.3 100 99 100 100 100 1_1998.5 100 98 100 100 100 1_20 100 100 100 100 100 100 1_21 99.3 100 99100 100 100 1_22 98.5 97.1 99 100 100 100 1_23 99.3 100 99 100 100 1001_24 98.5 94.1 100 100 100 100 1_25 99.3 100 99 98.5 100 98 1_26 99.3100 99 100 100 100 1_27 98.5 94.1 100 100 100 100 1_28 100 100 100 98.5100 98 1_29 98.5 97.1 99 100 100 100 1_30 100 100 100 100 100 100 1_3198.5 97.1 99 100 100 100 1_32 99.3 100 99 98.5 100 98 1_33 98.5 97.1 99100 100 100 1_34 97.8 97.1 98 100 100 100 1_35 98.5 97.1 99 98.5 100 981_36 100 100 100 100 100 100 1_37 97.8 97.1 98 98.5 100 98 1_38 98.597.1 99 100 100 100 1_39 99.3 97.1 100 100 100 100 1_40 97.8 97.1 98 100100 100 1_41 99.3 100 99 100 100 100 1_42 98.5 100 98 98.5 100 98 1_43100 100 100 100 100 100 1_44 97.8 97.1 98 100 100 100 1_45 98.5 100 98100 100 100 1_46 98.5 100 98 100 100 100 1_47 100 100 100 98.5 100 981_48 99.3 100 99 100 100 100 1_49 97.8 97.1 98 98.5 100 98 1_50 100 100100 98.5 100 98 1_51 98.5 97.1 99 100 100 100 1_52 97.8 94.1 99 98.5 10098 1_53 98.5 100 98 100 100 100 1_54 100 100 100 98.5 93.8 100 1_55 99.397.1 100 100 100 100 1_56 100 100 100 98.5 100 98 1_57 99.3 100 99 100100 100 1_58 98.5 97.1 99 100 100 100 1_59 99.3 100 99 100 100 100 1_6099.3 100 99 100 100 100 1_61 99.3 100 99 98.5 100 98 1_62 99.3 100 99100 100 100 1_63 99.3 100 99 100 100 100 1_64 100 100 100 98.5 100 981_65 99.3 100 99 98.5 100 98 1_66 99.3 97.1 100 100 100 100 1_67 99.3100 99 98.5 100 98 1_68 100 100 100 98.5 100 98 1_69 98.5 100 98 98.5100 98 1_70 99.3 100 99 100 100 100 1_71 99.3 100 99 100 100 100 1_7299.3 100 99 100 100 100 1_73 97.8 97.1 98 100 100 100 1_74 98.5 97.1 9998.5 100 98 1_75 99.2 100 99 98.5 100 98 1_76 98.5 100 98 100 100 1001_77 99.3 100 99 98.5 100 98 1_78 99.3 100 99 98.5 100 98 1_79 99.3 10099 98.5 100 98 1_80 98.5 100 98 98.5 100 98 1_81 98.5 97.1 99 98.5 10098 1_82 99.3 100 99 100 100 100 1_83 99.3 100 99 98.5 100 98 1_84 98.597.1 99 100 100 100 1_85 98.5 97.1 99 100 100 100 1_86 99.3 100 99 100100 100 1_87 99.3 97.1 100 100 100 100 1_88 98.5 100 98 100 100 100 1_8999.3 100 99 100 100 100 1_90 100 100 100 100 100 100 1_91 99.3 100 99100 100 100 1_92 99.3 100 99 100 100 100 1_93 99.3 100 99 98.5 100 981_94 98.5 97.1 99 100 100 100 1_95 99.3 100 99 100 100 100 1_96 99.3 10099 100 100 100 1_97 99.3 100 99 98.5 100 98 1_98 99.3 100 99 98.5 100 981_99 98.5 100 98 100 100 100 1_100 97 97.1 97 100 100 100 1_101 98.597.1 99 100 100 100 1_102 99.3 100 99 98.5 100 98 1_103 97.8 97.1 9898.5 100 98 1_104 100 100 100 100 100 100 1_105 98.5 100 98 100 100 1001_106 98.5 97.1 99 98.5 100 98 1_107 99.3 100 99 100 100 100 1_108 97.897.1 98 100 100 100 1_109 98.5 97.1 99 100 100 100 1_110 98.5 97.1 99100 100 100 1_111 98.5 100 98 100 100 100 1_112 98.5 97.1 99 100 100 1001_113 99.3 97.1 100 98.5 93.8 100 1_114 97.8 97.1 98 98.5 100 98 1_11599.3 100 99 100 100 100 1_116 100 100 100 100 100 100 1_117 97.8 94.1 9998.5 100 98 1_118 99.3 100 99 98.5 100 98 1_119 99.3 100 99 100 100 1001_120 98.5 100 98 97 93.8 98 1_121 99.3 97.1 100 100 100 100 1_122 98.5100 98 98.5 100 98 1_123 97.8 97.1 98 100 100 100 1_124 98.5 100 98 100100 100 1_125 98.5 97.1 99 98.5 93.8 100 1_126 99.3 100 99 100 100 1001_127 99.3 100 99 100 100 100 1_128 99.3 100 99 98.5 100 98 1_129 99.3100 99 100 100 100 1_130 97.8 97.1 98 100 100 100 1_131 97 94.1 98 100100 100 1_132 97.8 97.1 98 98.5 100 98 1_133 99.3 100 99 100 100 1001_134 99.3 100 99 100 100 100 1_135 98.5 97.1 99 100 100 100 1_136 97.8100 97 100 100 100 1_137 99.3 100 99 100 100 100 1_138 99.3 100 99 100100 100 1_139 98.5 97.1 99 97 100 96 1_140 98.5 94.1 100 100 100 1001_141 99.3 100 99 100 100 100 1_142 98.5 100 98 100 100 100 1_143 98.597.1 99 100 100 100 1_144 99.3 100 99 100 100 100 1_145 97.8 97.1 98 100100 100 1_146 100 100 100 100 100 100 1_147 99.3 100 99 100 100 1001_148 99.3 100 99 100 100 100 1_149 98.5 97.1 99 98.5 100 98 1_150 99.3100 99 100 100 100 1_151 99.3 100 99 100 100 100 1_152 98.5 97.1 99 100100 100 1_153 99.3 100 99 100 100 100 1_154 99.3 100 99 100 100 1001_155 98.5 100 98 100 100 100 1_156 100 100 100 100 100 100 1_157 97.897.1 98 98.5 100 98 1_158 98.5 97.1 99 100 100 100 1_159 97.8 97.1 98100 100 100 1_160 99.3 100 99 98.5 100 98 1_161 98.5 100 98 100 100 1001_162 99.3 100 99 100 100 100 1_163 97.8 100 97 100 100 100 1_164 99.3100 99 100 100 100 1_165 98.5 97.1 99 100 100 100 1_166 99.3 100 99 98.5100 98 1_167 99.3 100 99 100 100 100 1_168 99.2 100 99 100 100 100 1_16999.3 100 99 100 100 100 1_170 99.3 100 99 100 100 100 1_171 97.8 100 97100 100 100 1_172 98.5 97.1 99 98.5 100 98 1_173 99.3 100 99 100 100 1001_174 99.3 100 99 100 100 100 1_175 98.5 97.1 99 100 100 100 1_176 100100 100 98.5 100 98 1_177 98.5 97.1 99 100 100 100 1_178 99.3 100 9998.5 93.8 100 1_179 99.3 100 99 98.5 100 98 1_180 99.3 100 99 100 100100

TABLE 7 Training cohort Validation cohort Accuracy SensitivitySpecificity Accuracy Sensitivity Specificity SEQ ID NO: (%) (%) (%) (%)(%) (%) 5_6 98.5 97.1 99.0 93.9 87.5 96.0 5_11 98.5 97.1 99.0 97.0 87.5100 5_38 97.0 97.1 97.0 95.5 87.5 98.0 15_16 93.3 82.4 97.0 92.4 75.098.0 15_21 97.8 97.1 98.0 95.5 93.8 96.0 15_64 91.0 70.6 98.0 90.9 68.898.0 24_25 97.8 94.1 99.0 95.5 81.2 100 24_30 96.3 91.2 98.0 89.4 75.094.0 24_32 90.3 70.6 97.0 90.9 68.8 98.0 2_32 97.0 88.2 100 100 100 10032_36 94.8 82.4 99.0 89.4 68.8 96.0 15_32 92.5 76.5 98.0 95.5 87.5 98.03_38 97.0 97.1 97.0 97.0 100 96.0 38_39 93.3 82.4 97.0 87.9 75.0 92.038_64 87.3 61.8 96.0 87.9 62.5 96.0 3_45 96.3 85.3 100 97.0 100 96.045_58 96.3 91.2 98.0 83.3 75.0 86.0 45_64 95.5 94.1 96.0 95.5 87.5 98.02_55 96.3 88.2 99.0 100 100 100 6_55 95.5 85.3 99.0 90.9 81.2 94.0 55_6488.1 61.8 97.0 84.8 56.2 94.0 2_64 97.0 91.2 99.0 100 100 100 4_64 94.885.3 98.0 97.0 87.5 100 2_96 97.8 94.1 99.0 98.5 100 98.0 7_96 98.5 10098.0 93.9 93.8 94.0 96_97 85.1 61.8 93.0 77.3 31.2 92.0 2_97 96.3 88.299.0 100 100 100 3_97 98.5 97.1 99.0 98.5 100 98.0 5_97 96.3 91.2 98.097.0 93.8 98.0 2_162 96.3 88.2 99.0 98.5 100 98.0 3_162 97.8 94.1 99.0100 100 100 5_162 97.8 94.1 99.0 98.5 93.8 100

Example 3

<Selection of Gene Marker Using all Samples and Method for EvaluatingColorectal Cancer Discriminant Performance of Acquired Gene Marker>

In this Example, the samples in the training cohort and the validationcohort used in Examples 1 and 2 were integrated, and selection of a genemarker and evaluation of its colorectal cancer discriminant performancewere conducted using all of the samples.

Specifically, the miRNA expression levels in the serum of the 50colorectal cancer patients and the 150 healthy subjects obtained in thepreceding Reference Examples were normalized by quantile normalization.In order to acquire diagnostic markers with higher reliability, onlygenes having a gene expression level of 2⁶ or higher in 50% or more ofthe samples in either of the colorectal cancer patient group or thehealthy subject group were selected in the gene marker selection. Inorder to further acquire statistical significance for discriminating acolorectal cancer patient group from a healthy subject group, the Pvalue obtained by two-tailed t-test assuming equal variance as to eachgene expression level was corrected by the Bonfenoni method, and genesthat satisfied p<0.01 were selected as gene markers for use inexplanatory variables of a discriminant, and the obtained genes aredescribed in Table 8. In this way, hsa-miR-4697-5p, hsa-miR-3197,hsa-miR-675-5p, hsa-miR-4486, hsa-miR-7107-5p, hsa-miR-23a-3p,hsa-miR-4667-5p, hsa-miR-451a, hsa-miR-3940-5p, hsa-miR-8059,hsa-miR-6813-5p, hsa-miR-4492, hsa-miR-4476 and hsa-miR-6090 genes, andthe nucleotide sequences of SEQ ID NOs: 181 to 194 related thereto werefound in addition to the genes described in Table 2. As with thenucleotide sequences of SEQ ID NOs: 1 to 180, the results obtained aboutthe polynucleotides shown in SEQ ID NOs: 181 to 194 also showed that thegene measurement values were significantly lower (−) or higher (+) inthe colorectal cancer patient group than in the healthy subject group(Table 8). These results were able to be validated in the validationcohort. Thus, the presence or absence of colorectal cancer in the newlyobtained samples can be determined by the methods described in Examples1 and 2 by using the gene expression level measurement values describedin Table 8 either alone or in combination with the gene expression levelmeasurement values described in Table 2.

TABLE 8 Expression level in P value after colorectal cancer Bonferronipatient with respect SEQ ID NO: Gene name correction to healthy subject1 hsa-miR-6726-5p 5.31.E−62 − 2 hsa-miR-4257 1.09.E−61 − 3hsa-miR-6787-5p 2.44.E−47 − 4 hsa-miR-6780b-5p 2.11.E−42 + 5hsa-miR-3131 4.30.E−42 − 6 hsa-miR-7108-5p 3.00.E−35 + 7 hsa-miR-1343-3p4.27.E−43 − 8 hsa-miR-1247-3p 9.79.E−35 + 9 hsa-miR-4651 9.99.E−39 − 10hsa-miR-6757-5p 2.24.E−34 − 11 hsa-miR-3679-5p 3.50.E−37 + 12hsa-miR-7641 5.56.E−34 − 13 hsa-miR-6746-5p 1.02.E−31 − 14 hsa-miR-80721.54.E−27 + 15 hsa-miR-6741-5p 2.21.E−31 − 16 hsa-miR-1908-5p4.52.E−29 + 17 hsa-miR-6857-5p 3.92.E−22 + 18 hsa-miR-4746-3p3.57.E−31 + 19 hsa-miR-744-5p 7.34.E−32 + 20 hsa-miR-4792 1.24.E−27 + 21hsa-miR-564 2.13.E−30 − 22 hsa-miR-6791-5p 2.90.E−27 + 23hsa-miR-6825-5p 4.61.E−29 + 24 hsa-miR-6826-5p 2.05.E−29 − 25hsa-miR-4665-3p 7.74.E−29 + 26 hsa-miR-4467 5.07.E−27 + 27 hsa-miR-31885.96.E−29 + 28 hsa-miR-6125 2.14.E−23 + 29 hsa-miR-6756-5p 2.14.E−22 −30 hsa-miR-1228-3p 7.24.E−25 + 31 hsa-miR-8063 1.63.E−24 − 32hsa-miR-8069 9.97.E−22 + 33 hsa-miR-6875-5p 6.41.E−21 + 34 hsa-miR-31851.30.E−24 + 35 hsa-miR-4433b-3p 2.47.E−20 + 36 hsa-miR-6887-5p 5.17.E−26− 37 hsa-miR-128-1-5p 3.06.E−18 + 38 hsa-miR-6724-5p 4.44.E−21 + 39hsa-miR-1914-3p 2.19.E−16 − 40 hsa-miR-1225-5p 9.96.E−22 + 41hsa-miR-4419b 2.99.E−22 − 42 hsa-miR-7110-5p 1.00.E−22 + 43hsa-miR-187-5p 1.62.E−19 − 44 hsa-miR-3184-5p 2.98.E−20 + 45hsa-miR-204-3p 1.12.E−17 − 46 hsa-miR-5572 5.88.E−21 + 47hsa-miR-6729-5p 6.07.E−18 + 48 hsa-miR-615-5p 3.71.E−19 − 49hsa-miR-6749-5p 1.52.E−19 − 50 hsa-miR-6515-3p 1.14.E−15 + 51hsa-miR-3937 1.06.E−20 + 52 hsa-miR-6840-3p 3.27.E−16 − 53hsa-miR-6893-5p 3.70.E−20 − 54 hsa-miR-4728-5p 1.49.E−16 − 55hsa-miR-6717-5p 5.86.E−21 − 56 hsa-miR-7113-3p 1.99.E−19 + 57hsa-miR-4665-5p 4.71.E−16 − 58 hsa-miR-642b-3p 1.28.E−15 − 59hsa-miR-7109-5p 6.89.E−19 − 60 hsa-miR-6842-5p 5.06.E−19 + 61hsa-miR-4442 9.22.E−16 − 62 hsa-miR-4433-3p 2.94.E−16 + 63hsa-miR-4707-5p 1.21.E−7 + 64 hsa-miR-6126 3.89.E−16 + 65 hsa-miR-44493.16.E−20 + 66 hsa-miR-4706 1.73.E−16 − 67 hsa-miR-1913 3.48.E−16 + 68hsa-miR-602 1.60.E−16 + 69 hsa-miR-939-5p 4.02.E−16 + 70 hsa-miR-4695-5p2.61.E−14 + 71 hsa-miR-711 1.79.E−16 + 72 hsa-miR-6816-5p 5.98.E−14 + 73hsa-miR-4632-5p 4.56.E−14 + 74 hsa-miR-6721-5p 5.64.E−13 + 75hsa-miR-7847-3p 7.52.E−17 − 76 hsa-miR-6132 6.77.E−16 + 77hsa-miR-887-3p 3.26.E−14 + 78 hsa-miR-3679-3p 5.22.E−14 + 79hsa-miR-6784-5p 6.38.E−13 + 80 hsa-miR-1249 1.62.E−14 + 81hsa-miR-937-5p 8.71.E−13 − 82 hsa-miR-5195-3p 2.51.E−14 − 83hsa-miR-6732-5p 2.71.E−13 + 84 hsa-miR-4417 4.13.E−15 + 85 hsa-miR-42811.09.E−13 − 86 hsa-miR-4734 7.65.E−15 + 87 hsa-miR-6766-3p 1.32.E−13 +88 hsa-miR-663a 1.12.E−14 + 90 hsa-miR-6781-5p 1.88.E−11 + 91hsa-miR-1227-5p 6.26.E−12 + 92 hsa-miR-6845-5p 1.06.E−14 + 93hsa-miR-6798-5p 2.72.E−08 + 94 hsa-miR-3620-5p 7.80.E−10 + 95hsa-miR-1915-5p 1.02.E−11 − 96 hsa-miR-4294 1.22.E−12 − 97hsa-miR-642a-3p 5.69.E−12 − 98 hsa-miR-371a-5p 2.55.E−09 − 99hsa-miR-940 2.85.E−14 + 100 hsa-miR-4450 2.15.E−13 − 101 hsa-miR-4723-5p8.73.E−13 − 102 hsa-miR-1469 5.67.E−12 + 103 hsa-miR-6861-5p 2.03.E−12 −104 hsa-miR-7975 1.02.E−09 − 105 hsa-miR-6879-5p 6.99.E−11 + 106hsa-miR-6802-5p 1.21.E−10 − 107 hsa-miR-1268b 8.63.E−11 + 108hsa-miR-663b 1.02.E−10 − 109 hsa-miR-125a-3p 1.21.E−12 − 110hsa-miR-2861 4.18.E−13 − 111 hsa-miR-6088 6.31.E−12 − 112hsa-miR-4758-5p 1.17.E−10 − 113 hsa-miR-296-3p 1.20.E−08 − 114hsa-miR-6738-5p 1.29.E−09 − 115 hsa-miR-671-5 p 8.62.E−11 − 116hsa-miR-4454 4.34.E−10 − 117 hsa-miR-4516 3.61.E−10 − 118hsa-miR-7845-5p 7.69.E−09 + 119 hsa-miR-4741 2.27.E−09 + 120hsa-miR-92b-5p 2.68.E−09 + 121 hsa-miR-6795-5p 1.14.E−09 - 122hsa-miR-6805-3p 1.59.E−11 + 123 hsa-miR-4725-3p 6.13.E−07 + 124hsa-miR-6782-5p 1.59.E−08 + 125 hsa-miR-4688 5.22.E−07 − 126hsa-miR-6850-5p 7.32.E−08 + 127 hsa-miR-6777-5p 7.19.E−11 − 128hsa-miR-6785-5p 1.41.E−07 − 129 hsa-miR-7106-5p 6.63.E−09 − 130hsa-miR-3663-3p 3.69.E−09 − 131 hsa-miR-6131 1.40.E−09 − 132hsa-miR-1915-3p 6.80.E−08 + 133 hsa-miR-4532 2.71.E−07 − 134hsa-miR-6820-5p 1.32.E−07 − 135 hsa-miR-4689 3.51.E−09 − 136hsa-miR-4638-5p 2.60.E−07 − 137 hsa-miR-3656 1.23.E−07 + 138hsa-miR-3621 6.72.E−07 − 139 hsa-miR-6769b-5p 7.12.E−08 − 140hsa-miR-149-3p 1.99.E−07 − 141 hsa-miR-23b-3p 1.65.E−07 − 142hsa-miR-3135b 1.27.E−07 − 143 hsa-miR-6848-5p 3.54.E−06 + 144hsa-miR-6769a-5p 5.27.E−08 − 145 hsa-miR-4327 4.27.E−06 + 146hsa-miR-6765-3p 2.60.E−07 − 147 hsa-miR-6716-5p 1.00.E−06 + 148hsa-miR-6877-5p 1.64.E−06 − 149 hsa-miR-6727-5p 3.79.E−06 − 150hsa-miR-4534 4.38.E−06 − 151 hsa-miR-614 2.94.E−06 − 152 hsa-miR-12023.36.E−07 − 153 hsa-miR-575 5.28.E−08 − 154 hsa-miR-6870-5p 3.19.E−08 +155 hsa-miR-6722-3p 8.34.E−06 + 156 hsa-miR-7977 6.56.E−05 − 157hsa-miR-4649-5p 1.23.E−05 − 158 hsa-miR-4675 3.15.E−07 − 159hsa-miR-6075 6.53.E−05 + 160 hsa-miR-6779-5p 5.68.E−07 − 161hsa-miR-4271 1.02.E−05 − 162 hsa-miR-3196 2.40.E−06 + 163hsa-miR-6803-5p 3.32.E−03 + 164 hsa-miR-6789-5p 1.02.E−06 + 165hsa-miR-4648 7.63.E−08 + 167 hsa-miR-4749-5p 3.78.E−05 + 168hsa-miR-4505 7.82.E−05 + 169 hsa-miR-5698 2.28.E−04 − 170hsa-miR-1199-5p 2.58.E−04 − 171 hsa-miR-4763-3p 1.20.E−03 + 172hsa-miR-1231 2.42.E−35 + 173 hsa-miR-1233-5p 4.01.E−32 − 174hsa-miR-150-3p 4.05.E−09 − 175 hsa-miR-1225-3p 3.42.E−13 + 176hsa-miR-92a-2-5p 3.89.E−08 + 177 hsa-miR-423-5p 1.73.E−06 − 178hsa-miR-1268a 2.52.E−05 + 179 hsa-miR-128-2-5p 5.33.E−06 − 180hsa-miR-24-3p 1.01.E−07 − 181 hsa-miR-4697-5p 4.79.E−05 − 182hsa-miR-3197 1.62.E−04 + 183 hsa-miR-675-5p 2.19.E−04 − 184 hsa-miR-44864.27.E−04 + 185 hsa-miR-7107-5p 4.72.E−04 − 186 hsa-miR-23a-3p 1.53.E−03− 187 hsa-miR-4667-5p 2.51.E−03 + 188 hsa-miR-451a 3.74.E−03 − 189hsa-miR-3940-5p 4.95.E−03 + 190 hsa-miR-8059 5.22.E−03 − 191hsa-miR-6813-5p 5.33.E−03 + 192 hsa-miR-4492 9.03.E−03 + 193hsa-miR-4476 9.04.E−03 − 194 hsa-miR-6090 9.46.E−03 +

Example 4

<Method for Evaluating Colorectal Cancer-Specific DiscriminantPerformance by Combination of Multiple Gene Markers Using Samples in theValidation Cohort>

In this Example, a gene for diagnosis is selected by comparing geneexpression levels of miRNAs in serum between colorectal cancer patientsand a control group that consist of healthy subjects, pancreatic cancerpatients, bile duct cancer patients, stomach cancer patients, esophagealcancer patients, liver cancer patients, and benign pancreaticobiliarydisease patients in the same way as the method described in Example 1,using the gene markers selected in Example 1, and targeting the trainingcohort as the sample group described in Reference Example 2. Thepolynucleotides consisting of the nucleotide sequences represented bySEQ ID NOs: 606 to 614 thus selected were further combined therewith tostudy a method for evaluating colorectal cancer-specific discriminantperformance.

Specifically, first, the miRNA expression levels in the training cohortand the validation cohort obtained in Reference Example 2 were combinedand normalized by quantile normalization. Next, Fisher's discriminantanalysis was conducted as to combinations of 1 to 6 expression levelmeasurement values comprising at least one or more of the expressionlevel measurement values of the polynucleotides consisting of thenucleotide sequences represented by SEQ ID NOs: 1 to 171 and 606 to 614,to construct a discriminant for determining the presence or absence ofcolorectal cancer. Next, accuracy, sensitivity, and specificity in thevalidation cohort were calculated using the discriminant thus prepared,with the colorectal cancer patient group as a positive sample group and,on the other hand, the healthy subject group, the pancreatic cancerpatient group, the bile duct cancer patient group, the stomach cancerpatient group, the esophageal cancer patient group, the liver cancerpatient group, and the benign pancreaticobiliary disease patient groupas a negative sample group. The discriminant performance of the selectedpolynucleotides was validated using independent samples.

Most of polynucleotides consisting of the nucleotide sequencesrepresented by these SEQ ID NOs (SEQ ID NOs: 1 to 194 and 606 to 614corresponding to the miRNA markers of Table 1) or complementarysequences thereof were able to provide relatively high accuracy,sensitivity, and specificity in the determination of the presence orabsence of colorectal cancer, and furthermore, were able to specificallydiscriminate colorectal cancer from the other cancers. For example,among the combinations of multiple polynucleotides selected from thegroup consisting of polynucleotides consisting of the nucleotidesequences represented by SEQ ID NOs: 5, 13, 15, 24, 32, 38, 41, 45, 55,57, 64, 72, 75, 77, 96, 97, 115, 162, 163, 173, 189, 606, 607, 608, 609,610, 611, 612, 613 and 614, or complementary sequences thereof (thecancer type-specific polynucleotide group 1) as polynucleotides capableof specifically binding to target markers, combinations comprising atleast one or more polynucleotide(s) selected from the group consistingof polynucleotides consisting of the nucleotide sequences represented bySEQ ID NOs: 5, 45, 57, 96, and 606, or complementary sequences thereof(the cancer type-specific polynucleotide group 2) included in the cancertype-specific polynucleotide group 1 were able to specificallydiscriminate colorectal cancer from the other cancers with highaccuracy.

The number of the aforementioned polynucleotides with cancer typespecificity in the combination can be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 ormore for the combination. The combinations of 6 or more of thesepolynucleotides were able to exhibit discriminant accuracy of 90% orhigher.

Specifically, the discriminant accuracy of the measurement using thepolynucleotide that consists of the nucleotide sequence represented bySEQ ID NO: 5 or a complementary sequence thereof is shown in Table 9-1.The measurement using the combination of one polynucleotide comprisingat least one polynucleotide consisting of the nucleotide sequencerepresented by SEQ ID NO: 5 or a complementary sequence thereofexhibited the highest accuracy of 90.1% in the training cohort andaccuracy of 87.6% in the validation cohort. Also, for example, themeasurement using the combinations of two polynucleotides comprising atleast one polynucleotide consisting of the nucleotide sequencerepresented by SEQ ID NO: 5 or a complementary sequence thereofexhibited the highest accuracy of 91.7% in the training cohort andaccuracy of 88.8% in the validation cohort. Furthermore, for example,the measurement using the combinations of three polynucleotidescomprising at least one polynucleotide consisting of the nucleotidesequence represented by SEQ ID NO: 5 or a complementary sequence thereofexhibited the highest accuracy of 94.0% in the training cohort andaccuracy of 91.2% in the validation cohort. Furthermore, for example,the measurement using the combinations of four polynucleotidescomprising at least one polynucleotide consisting of the nucleotidesequence represented by SEQ ID NO: 5 or a complementary sequence thereofexhibited the highest accuracy of 95.6% in the training cohort andaccuracy of 93.6% in the validation cohort. Furthermore, for example,the measurement using the combinations of five polynucleotidescomprising at least one polynucleotide consisting of the nucleotidesequence represented by SEQ ID NO: 5 or a complementary sequence thereofexhibited the highest accuracy of 96.4% in the training cohort andaccuracy of 94.8% in the validation cohort. Furthermore, for example,the measurement using the combinations of six polynucleotides comprisingat least one polynucleotide consisting of the nucleotide sequencerepresented by SEQ ID NO: 5 or a complementary sequence thereofexhibited the highest accuracy of 96.9% in the training cohort andaccuracy of 94.7% in the validation cohort.

The discriminant accuracy of the measurement using the polynucleotideconsisting of the nucleotide sequence represented by SEQ ID NO: 45 or acomplementary sequence thereof is shown in Table 9-2. The measurementusing the combination of one polynucleotide comprising at least onepolynucleotide consisting of the nucleotide sequence represented by SEQID NO: 45 or a complementary sequence thereof exhibited the highestaccuracy of 56.7% in the training cohort and accuracy of 55.4% in thevalidation cohort. Also, for example, the measurement using thecombinations of two polynucleotides comprising at least onepolynucleotide consisting of the nucleotide sequence represented by SEQID NO: 45 or a complementary sequence thereof exhibited the highestaccuracy of 90.7% in the training cohort and accuracy of 88.4% in thevalidation cohort. Furthermore, for example, the measurement using thecombinations of three polynucleotides comprising at least onepolynucleotide consisting of the nucleotide sequence represented by SEQID NO: 45 or a complementary sequence thereof exhibited the highestaccuracy of 94.0% in the training cohort and accuracy of 89.6% in thevalidation cohort. Furthermore, for example, the measurement using thecombinations of four polynucleotides comprising at least onepolynucleotide consisting of the nucleotide sequence represented by SEQID NO: 45 or a complementary sequence thereof exhibited the highestaccuracy of 95.2% in the training cohort and accuracy of 91.6% in thevalidation cohort. Furthermore, for example, the measurement using thecombinations of five polynucleotides comprising at least onepolynucleotide consisting of the nucleotide sequence represented by SEQID NO: 45 or a complementary sequence thereof exhibited the highestaccuracy of 96.4% in the training cohort and accuracy of 94.4% in thevalidation cohort. Furthermore, for example, the measurement using thecombinations of six polynucleotides comprising at least onepolynucleotide consisting of the nucleotide sequence represented by SEQID NO: 45 or a complementary sequence thereof exhibited the highestaccuracy of 97.6% in the training cohort and accuracy of 92.6% in thevalidation cohort.

The discriminant accuracy of the measurement using the polynucleotideconsisting of the nucleotide sequence represented by SEQ ID NO: 57 or acomplementary sequence thereof is shown in Table 9-3. The measurementusing the combination of one polynucleotide comprising at least onepolynucleotide consisting of the nucleotide sequence represented by SEQID NO: 57 or a complementary sequence thereof exhibited the highestaccuracy of 60.2% in the training cohort and accuracy of 60.6% in thevalidation cohort. Also, for example, the measurement using thecombinations of two polynucleotides comprising at least onepolynucleotide consisting of the nucleotide sequence represented by SEQID NO: 57 or a complementary sequence thereof exhibited the highestaccuracy of 86.7% in the training cohort and accuracy of 83.7% in thevalidation cohort. Furthermore, for example, the measurement using thecombinations of three polynucleotides comprising at least onepolynucleotide consisting of the nucleotide sequence represented by SEQID NO: 57 or a complementary sequence thereof exhibited the highestaccuracy of 92.4% in the training cohort and accuracy of 90.0% in thevalidation cohort. Furthermore, for example, the measurement using thecombinations of four polynucleotides comprising at least onepolynucleotide consisting of the nucleotide sequence represented by SEQID NO: 57 or a complementary sequence thereof exhibited the highestaccuracy of 95.2% in the training cohort and accuracy of 91.2% in thevalidation cohort. Furthermore, for example, the measurement using thecombinations of five polynucleotides comprising at least onepolynucleotide consisting of the nucleotide sequence represented by SEQID NO: 57 or a complementary sequence thereof exhibited the highestaccuracy of 96.2% in the training cohort and accuracy of 94.8% in thevalidation cohort. Furthermore, for example, the measurement using thecombinations of six polynucleotides comprising at least onepolynucleotide consisting of the nucleotide sequence represented by SEQID NO: 57 or a complementary sequence thereof exhibited the highestaccuracy of 96.9% in the training cohort and accuracy of 93.6% in thevalidation cohort.

The discriminant accuracy of the measurement using the polynucleotideconsisting of the nucleotide sequence represented by SEQ ID NO: 96 or acomplementary sequence thereof is shown in Table 9-4. The measurementusing the combination of one polynucleotide comprising at least onepolynucleotide consisting of the nucleotide sequence represented by SEQID NO: 96 or a complementary sequence thereof exhibited the highestaccuracy of 57.9% in the training cohort and accuracy of 59.4% in thevalidation cohort. Also, for example, the measurement using thecombinations of two polynucleotides comprising at least onepolynucleotide consisting of the nucleotide sequence represented by SEQID NO: 96 or a complementary sequence thereof exhibited the highestaccuracy of 85.9% in the training cohort and accuracy of 83.7% in thevalidation cohort. Furthermore, for example, the measurement using thecombinations of three polynucleotides comprising at least onepolynucleotide consisting of the nucleotide sequence represented by SEQID NO: 96 or a complementary sequence thereof exhibited the highestaccuracy of 92.6% in the training cohort and accuracy of 90.4% in thevalidation cohort. Furthermore, for example, the measurement using thecombinations of four polynucleotides comprising at least onepolynucleotide consisting of the nucleotide sequence represented by SEQID NO: 96 or a complementary sequence thereof exhibited the highestaccuracy of 94.4% in the training cohort and accuracy of 91.2% in thevalidation cohort. Furthermore, for example, the measurement using thecombinations of five polynucleotides comprising at least onepolynucleotide consisting of the nucleotide sequence represented by SEQID NO: 96 or a complementary sequence thereof exhibited the highestaccuracy of 96.0% in the training cohort and accuracy of 94.0% in thevalidation cohort. Furthermore, for example, the measurement using thecombinations of six polynucleotides comprising at least onepolynucleotide consisting of the nucleotide sequence represented by SEQID NO: 96 or a complementary sequence thereof exhibited the highestaccuracy of 96.3% in the training cohort and accuracy of 93.6% in thevalidation cohort.

The discriminant accuracy of the measurement using the polynucleotideconsisting of the nucleotide sequence represented by SEQ ID NO: 606 or acomplementary sequence thereof is shown in Table 9-5. The measurementusing the combination of one polynucleotide comprising at least onepolynucleotide consisting of the nucleotide sequence represented by SEQID NO: 606 or a complementary sequence thereof exhibited the highestaccuracy of 59.4% in the training cohort and accuracy of 58.6% in thevalidation cohort. Also, for example, the measurement using thecombinations of two polynucleotides comprising at least onepolynucleotide consisting of the nucleotide sequence represented by SEQID NO: 606 or a complementary sequence thereof exhibited the highestaccuracy of 86.6% in the training cohort and accuracy of 82.9% in thevalidation cohort. Furthermore, for example, the measurement using thecombinations of three polynucleotides comprising at least onepolynucleotide consisting of the nucleotide sequence represented by SEQID NO: 606 or a complementary sequence thereof exhibited the highestaccuracy of 92.6% in the training cohort and accuracy of 91.2% in thevalidation cohort. Furthermore, for example, the measurement using thecombinations of four polynucleotides comprising at least onepolynucleotide consisting of the nucleotide sequence represented by SEQID NO: 606 or a complementary sequence thereof exhibited the highestaccuracy of 94.8% in the training cohort and accuracy of 90.0% in thevalidation cohort. Furthermore, for example, the measurement using thecombinations of five polynucleotides comprising at least onepolynucleotide consisting of the nucleotide sequence represented by SEQID NO: 606 or a complementary sequence thereof exhibited the highestaccuracy of 96.0% in the training cohort and accuracy of 93.6% in thevalidation cohort. Furthermore, for example, the measurement using thecombinations of six polynucleotides comprising at least onepolynucleotide consisting of the nucleotide sequence represented by SEQID NO: 606 or a complementary sequence thereof exhibited the highestaccuracy of 95.3% in the training cohort and accuracy of 93.6% in thevalidation cohort.

The expression level measurement values of the nucleotide sequencesrepresented by SEQ ID NOs: 5, 45, 57, 75, 162, and 607 were comparedamong 34 colorectal cancer patients, 103 healthy subject, 69 pancreaticcancer patients, 66 bile duct cancer patients, 30 stomach cancerpatients, 33 esophageal cancer patients, 32 liver cancer patients, and15 benign pancreaticobiliary disease patients in the training cohort. Asa result, a scatter diagram that significantly separated thediscriminant score of the colorectal cancer patient group from thediscriminant scores of the other groups was obtained in the trainingcohort (see the upper diagram of FIG. 4 ). These results were alsoreproducible in the validation cohort (see the lower diagram of FIG. 4).

TABLE 9-1 Training cohort Validation cohort Accuracy SensitivitySpecificity Accuracy Sensitivity Specificity SEQ ID NO: (%) (%) (%) (%)(%) (%) 5 90.1 100 89.3 87.6 87.5 87.7 5_608 91.7 91.2 91.7 88.8 62.590.6 5_45_607 94 91.2 94.2 91.2 75 92.3 5_45_57_607 95.6 88.2 96.2 93.662.5 95.7 5_45_57_75_607 96.3 84.8 97.4 93.1 62.5 95.9 5_45_96_606_60796.4 97.1 96.4 94.8 87.5 95.3 5_45_57_97_115_607 96.9 88.2 97.7 94.775.0 96.5 5_45_57_97_162_607 96.9 88.2 97.7 94.1 68.8 96.55_45_57_162_607_613 96.9 88.2 97.7 94.1 62.5 97.1 5_45_57_97_607_61296.9 94.1 97.1 94.1 81.2 95.3 5_13_45_57_606_607 96.9 91.2 97.4 93.668.8 95.9 5_45_96_189_606_608 95.3 94.1 95.4 94.7 75 96.55_45_57_96_189_606 96.3 97.1 96.3 93.6 75 95.3 5_24_45_57_96_608 95.394.1 95.4 92.6 56.2 95.9 5_45_57_162_607_610 95.8 85.3 96.8 93.6 62.596.5 5_45_57_189_606_607 96.1 91.2 96.6 93.6 75 95.3

TABLE 9-2 Training cohort Validation cohort Accuracy SensitivitySpecificity Accuracy Sensitivity Specificity SEQ ID NO: (%) (%) (%) (%)(%) (%) 45 56.7 61.8 56.3 55.4 56.2 55.3 5_45 90.7 100 90 88.4 87.5 88.55_45_57 94 94.1 94 89.6 81.2 90.2 5_45_57_97 95.2 94.1 95.3 91.6 81.292.3 5_45_96_606_607 95.5 91.2 96.0 95.2 87.5 95.9 5_45_57_75_607 96.487.9 97 94.4 62.5 96.6 5_45_57_75_606_607 97.6 87.9 98.6 92.6 62.5 95.35_45_57_77_607_613 97.4 94.1 97.7 94.1 75.0 95.9 5_45_57_97_606_607 97.194.1 97.4 94.1 81.2 95.3 5_45_57_75_77_607 97.1 90.9 97.7 93.1 68.8 95.35_32_45_57_96_606 96.3 97.1 96.3 93.6 68.8 95.9 5_24_45_57_96_606 96.197.1 96 93.1 68.8 95.3 5_45_57_96_162_606 95.5 91.2 96 94.7 81.2 95.95_15_45_75_96_606 95.5 100 95.1 93.6 81.2 94.8 5_32_45_57_162_607 95.885.3 96.8 93.6 62.5 96.5 38_45_96_606_608_611 87.1 88.2 87.0 86.2 68.887.8

TABLE 9-3 Training cohort Validation cohort Accuracy SensitivitySpecificity Accuracy Sensitivity Specificity SEQ ID NO: (%) (%) (%) (%)(%) (%) 57 60.2 70.6 59.5 60.6 56.2 60.9 24_57 86.7 91.2 86.4 83.7 62.585.1 5_57_608 92.4 88.2 92.8 90 68.8 91.5 5_45_57_608 95.2 91.2 95.591.2 62.5 93.2 24_41_57_45_96 94.5 94.1 94.5 88.8 56.2 91.95_45_57_607_612 96.2 94.1 96.4 94.8 68.8 96.6 5_45_57_606_607_608 96.991.2 97.4 93.6 68.8 95.9 5_13_45_57_75_607 96.9 90.9 97.4 93.1 68.8 95.35_45_57_64_75_607 96.9 90.9 97.4 92.6 68.8 94.8 5_45_55_57_607_613 96.991.2 97.4 92.6 68.8 94.8 5_45_55_57_75_607 96.6 87.9 97.4 92.6 68.8 94.85_38_45_57_96_607 96.3 88.2 97.1 94.1 68.8 96.5 5_45_57_75_162_607 96.687.9 97.4 94.1 62.5 97.1 5_45_57_75_162_609 94.2 97 94 91.5 62.5 94.25_45_57_64_96_607 95.5 88.2 96.3 94.7 75 96.5 57_64_96_606_608_611 90.691.2 90.5 88.3 75.0 89.5

TABLE 9-4 Training cohort Validation cohort Accuracy SensitivitySpecificity Accuracy Sensitivity Specificity SEQ ID NO: (%) (%) (%) (%)(%) (%) 96 57.9 58.8 57.8 59.4 62.5 59.1 41_96 85.9 88.2 85.7 83.7 62.585.1 5_96_606 92.6 100 92.1 90.4 87.5 90.6 5_45_57_96 94.4 91.2 94.791.2 75 92.3 38_96_606_608_611 86.4 91.2 85.9 85.6 75 86.65_45_57_96_607 96 91.2 96.4 94 68.8 95.7 38_72_96_606_608_611 89.0 88.289.0 87.7 75.0 88.9 32_38_96_606_608_611 89.8 88.2 89.9 86.7 68.8 88.438_96_163_606_608_611 87.4 85.3 87.6 85.1 68.8 86.6 64_72_96_162_609_61181.9 85.3 81.6 81.8 81.2 81.9 38_64_96_163_606_608 87.4 91.2 87.1 86.768.8 88.4 5_45_57_75_96_606 96.3 93.9 96.6 93.6 81.2 94.85_15_45_57_96_606 95.5 91.2 96 94.1 87.5 94.8 5_41_45_57_96_606 94.891.2 95.1 94.1 87.5 94.8 5_41_45_96_189_606 94.5 100 94 93.1 75 94.85_45_75_96_189_606 94.8 97 94.5 94.7 75 96.5

TABLE 9-5 Training cohort Validation cohort 606 Accuracy SensitivitySpecificity Accuracy Sensitivity Specificity SEQ ID NO: (%) (%) (%) (%)(%) (%) 606 59.4 61.8 59.3 58.6 50 59.1 75_606 86.6 84.8 86.8 82.9 62.584.3 5_606_610 92.6 97.1 92.3 91.2 81.2 91.9 5_45_96_606 94.8 100 94.590 87.5 90.2 64_96_606_608_611 86.4 91.2 85.9 85.6 75.0 86.65_45_57_606_610 96 94.1 96.2 93.6 68.8 95.3 64_96_162_609_610_611 81.985.3 81.6 81.4 81.2 81.4 38_64_96_606_608_611 88.7 88.2 88.8 87.8 75.089.0 64_72_96_606_608_611 89.0 88.2 89.0 88.2 75.0 89.564_96_97_606_608_611 89.7 88.2 89.9 89.4 75.0 90.7 45_64_96_606_608_61189.8 88.2 89.9 88.8 75.0 90.1 5_24_45_96_189_606 95.3 100 94.8 93.6 62.596.5 5_15_45_96_189_606 94 94.1 94 94.1 75 95.9 5_45_96_189_606_613 9597.1 94.8 94.7 81.2 95.9 5_45_72_96_189_606 95 97.1 94.8 94.7 81.2 95.95_15_32_45_96_606 95.3 97.1 95.1 93.6 68.8 95.9

Comparative Example 1

<Colorectal Cancer Discriminant Performance of an Existing Tumor Markerin Blood>

The concentration of the existing tumor marker CEA in blood was measuredin the training cohort and the validation cohort obtained in thepreceding Reference Examples. When the concentration of the tumor markerin blood is higher than the reference value described in Non PatentLiterature 4 (CEA: 5 ng/mL), subjects are generally suspected of havingcancer. Thus, whether or not the concentration of CEA in blood exceededits reference value was confirmed for each sample, and the results wereassessed for the ability of the tumor marker to detect cancer incolorectal cancer patients. The sensitivity of the existing marker inthe training cohort and the validation cohort was calculated. Theresults are shown in Tables 5-1 and 5-2. The sensitivity of CEA was aslow as 26.5% in the training cohort and was as low as 43.8% in thevalidation cohort, demonstrating that the marker is not useful in thedetection of colorectal cancer (Tables 5-1 and 5-2).

On the other hand, as shown above in Tables 3 and 6 of Examples 1 and 2,it can be concluded that in all of the polynucleotides consisting of thenucleotide sequences represented by SEQ ID NOs: 1 to 180, combinationsof 1 or 2 polynucleotides exhibiting sensitivity beyond the existingcolorectal cancer marker are present, and thus such polynucleotidesserve as excellent diagnosis markers.

As shown in these Examples and Comparative Example, the kit, etc., andthe method of the present invention can detect colorectal cancer moresensitively than the existing tumor marker and therefore permit earlydetection and treatment of colorectal cancer. As a result, improvementin survival rate and a therapeutic option of endoscopic operation, whichplaces less burden on patients, can also be provided.

INDUSTRIAL APPLICABILITY

According to the present invention, colorectal cancer can be effectivelydetected by a simple and inexpensive method. This enables earlydetection, diagnosis and treatment of colorectal cancer. The method ofthe present invention can detect colorectal cancer with limitedinvasiveness using the blood of a patient and therefore allowscolorectal cancer to be detected conveniently and rapidly.

All publications, patents, and patent applications cited herein areincorporated herein by reference in their entirety.

1. A method for detecting colorectal cancer in a human subject,comprising: measuring an expression level of hsa-miR-1231 in a blood,serum, or plasma sample from the subject, comparing the measuredexpression level of hsa-miR-1231 to a control expression level for ahealthy subject; detecting an increased level of hsa-miR-1231 in thesample from the subject as compared to the control expression level fromthe sample from the healthy subject; wherein the increased level ofhsa-miR-1231 indicates that the subject has colorectal cancer; andwherein the method further comprises treating the subject for thecolorectal cancer or performing a diagnostic procedure on the subjectwith the colorectal cancer; wherein the treating comprises surgery,radiotherapy, chemotherapy or a combination thereof; and wherein thediagnostic procedure comprises fecal occult blood, rectal examination,colonoscopy, barium enema, CT, MRI, bone scintigraphy, or a combinationthereof.
 2. The method according to claim 1, wherein the expressionlevel of hsa-miR-1231 in the sample is measured by using a kitcomprising a nucleic acid(s) that specifically binds to hsa-miR-1231. 3.The method according to claim 2, wherein the kit further comprises atleast one nucleic acid capable of specifically binding to at least onepolynucleotide selected from the group consisting of other colorectalcancer markers: miR-4257, miR-6787-5p, miR-6780b-5p, miR-3131,miR-7108-5p, miR-1343-3p, miR-1247-3p, miR-4651, miR-6757-5p,miR-3679-5p, miR-7641, miR-6746-5p, miR-8072, miR-6741-5p, miR-1908-5p,miR-6857-5p, miR-4746-3p, miR-744-5p, miR-4792, miR-564, miR-6791-5p,miR-6825-5p, miR-6826-5p, miR-4665-3p, miR-4467, miR-3188, miR-6125,miR-6756-5p, miR-1228-3p, miR-8063, miR-8069, miR-6875-5p, miR-3185,miR-4433b-3p, miR-6887-5p, miR-128-1-5p, miR-6724-5p, miR-1914-3p,miR-1225-5p, miR-4419b, miR-7110-5p, miR-187-5p, miR-3184-5p,miR-204-3p, miR-5572, miR-6729-5p, miR-615-5p, miR-6749-5p, miR-6515-3p,miR-6840-3p, miR-6893-5p, miR-4728-5p, miR-6717-5p, miR-7113-3p,miR-4665-5p, miR-642b-3p, miR-7109-5p, miR-6842-5p, miR-4442,miR-4433-3p, miR-4707-5p, miR-6126, miR-4449, miR-4706, miR-1913,miR-602, miR-939-5p, miR-4695-5p, miR-711, miR-6816-5p, miR-4632-5p,miR-6721-5p, miR-7847-3p, miR-6132, miR-887-3p, miR-3679-3p,miR-6784-5p, miR-1249, miR-937-5p, miR-5195-3p, miR-6732-5p, miR-4417,miR-4281, miR-4734, miR-6766-3p, miR-663a, miR-4513, miR-6781-5p,miR-1227-5p, miR-6845-5p, miR-6798-5p, miR-3620-5p, miR-1915-5p,miR-4294, miR-642a-3p, miR-371a-5p, miR-940, miR-4450, miR-4723-5p,miR-1469, miR-6861-5p, miR-7975, miR-6879-5p, miR-6802-5p, miR-1268b,miR-663b, miR-125a-3p, miR-2861, miR-6088, miR-4758-5p, miR-296-3p,miR-6738-5p, miR-671-5p, miR-4454, miR-4516, miR-7845-5p, miR-4741,miR-92b-5p, miR-6795-5p, miR-6805-3p, miR-4725-3p, miR-6782-5p,miR-4688, miR-6850-5p, miR-6777-5p, miR-6785-5p, miR-7106-5p,miR-3663-3p, miR-6131, miR-1915-3p, miR-4532, miR-6820-5p, miR-4689,miR-4638-5p, miR-3656, miR-3621, miR-6769b-5p, miR-149-3p, miR-23b-3p,miR-3135b, miR-6848-5p, miR-6769a-5p, miR-4327, miR-6765-3p,miR-6716-5p, miR-6877-5p, miR-6727-5p, miR-4534, miR-614, miR-1202,miR-575, miR-6870-5p, miR-6722-3p, miR-7977, miR-4649-5p, miR-4675,miR-6075, miR-6T79-5p, miR-4271, miR-3196, miR-6803-5p, miR-6789-5p,miR-4648, miR-4508, miR-4749-5p, miR-4505, miR-5698, miR-1199-5p,miR-4763-3p, miR-6836-3p, miR-3195, miR-718, miR-3178, miR-638,miR-4497, miR-6085, miR-6752-5p and miR-135a-3p and/or miR-1233-5p,miR-150-3p, miR-1225-3p, miR-92a-2-5p, miR-423-5p, miR-1268a,miR-128-2-5p, miR-24-3p, miR-4697-5p, miR-3197, miR-675-5p, miR-4486,miR-7107-5p, miR-23a-3p, miR-4667-5p, miR-451a, miR-3940-5p, miR-8059,miR-6813-5p, miR-4492, miR-4476, and miR-6090.
 4. The method accordingto claim 1, comprising performing the diagnostic procedure on thesubject.
 5. The method according to claim 1, wherein the expressionlevel of hsa-miR-1231 in the sample is measured by using a devicecomprising a nucleic acid(s) that specifically binds to hsa-miR-1231. 6.The method according to claim 5, wherein the device further comprises atleast one nucleic acid capable of specifically binding to at least onepolynucleotide selected from the group consisting of other colorectalcancer markers: miR-4257, miR-6787-5p, miR-6780b-5p, miR-3131,miR-7108-5p, miR-1343-3p, miR-1247-3p, miR-4651, miR-6757-5p,miR-3679-5p, miR-7641, miR-6746-5p, miR-8072, miR-6741-5p, miR-1908-5p,miR-6857-5p, miR-4746-3p, miR-744-5p, miR-4792, miR-564, miR-6791-5p,miR-6825-5p, miR-6826-5p, miR-4665-3p, miR-4467, miR-3188, miR-6125,miR-6756-5p, miR-1228-3p, miR-8063, miR-8069, miR-6875-5p, miR-3185,miR-4433b-3p, miR-6887-5p, miR-128-1-5p, miR-6724-5p, miR-1914-3p,miR-1225-5p, miR-4419b, miR-7110-5p, miR-187-5p, miR-3184-5p,miR-204-3p, miR-5572, miR-6729-5p, miR-615-5p, miR-6749-5p, miR-6515-3p,miR-6840-3p, miR-6893-5p, miR-4728-5p, miR-6717-5p, miR-7113-3p,miR-4665-5p, miR-642b-3p, miR-7109-5p, miR-6842-5p, miR-4442,miR-4433-3p, miR-4707-5p, miR-6126, miR-4449, miR-4706, miR-1913,miR-602, miR-939-5p, miR-4695-5p, miR-711, miR-6816-5p, miR-4632-5p,miR-6721-5p, miR-7847-3p, miR-6132, miR-887-3p, miR-3679-3p,miR-6784-5p, miR-1249, miR-937-5p, miR-5195-3p, miR-6732-5p, miR-4417,miR-4281, miR-4734, miR-6766-3p, miR-663a, miR-4513, miR-6781-5p,miR-1227-5p, miR-6845-5p, miR-6798-5p, miR-3620-5p, miR-1915-5p,miR-4294, miR-642a-3p, miR-371a-5p, miR-940, miR-4450, miR-4723-5p,miR-1469, miR-6861-5p, miR-7975, miR-6879-5p, miR-6802-5p, miR-1268b,miR-663b, miR-125a-3p, miR-2861, miR-6088, miR-4758-5p, miR-296-3p,miR-6738-5p, miR-671-5p, miR-4454, miR-4516, miR-7845-5p, miR-4741,miR-92b-5p, miR-6795-5p, miR-6805-3p, miR-4725-3p, miR-6782-5p,miR-4688, miR-6850-5p, miR-6777-5p, miR-6785-5p, miR-7106-5p,miR-3663-3p, miR-6131, miR-1915-3p, miR-4532, miR-6820-5p, miR-4689,miR-4638-5p, miR-3656, miR-3621, miR-6769b-5p, miR-149-3p, miR-23b-3p,miR-3135b, miR-6848-5p, miR-6769a-5p, miR-4327, miR-6765-3p,miR-6716-5p, miR-6877-5p, miR-6727-5p, miR-4534, miR-614, miR-1202,miR-575, miR-6870-5p, miR-6722-3p, miR-7977, miR-4649-5p, miR-4675,miR-6075, miR-6779-5p, miR-4271, miR-3196, miR-6803-5p, miR-6789-5p,miR-4648, miR-4508, miR-4749-5p, miR-4505, miR-5698, miR-1199-5p,miR-4763-3p, miR-6836-3p, miR-3195, miR-718, miR-3178, miR-638,miR-4497, miR-6085, miR-6752-5p and miR-135a-3p and/or miR-1233-5p,miR-150-3p, miR-1225-3p, miR-92a-2-5p, miR-423-5p, miR-1268a,miR-128-2-5p, miR-24-3p, miR-4697-5p, miR-3197, miR-675-5p, miR-4486,miR-7107-5p, miR-23a-3p, miR-4667-5p, miR-451a, miR-3940-5p, miR-8059,miR-6813-5p, miR-4492, miR-4476, and miR-6090.